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Bethesda, MD 20892-2094, Probiotic blocks staph bacteria from colonizing people, Engineering skin grafts for complex body parts, Links found between viruses and neurodegenerative diseases, Bivalent boosters provide better protection against severe COVID-19. More recently, Myers and co-workers48, and others, have developed efficient algorithms for exploiting such linking information. The observed sequence identity in fourfold degenerate sites was 67%, and the estimated number of substitutions per site, between 0.46 and 0.47, was similar to that in the ancestral repeat sites (see Supplementary Information). For each mouse chromosome, its (G+C) content is depicted as a greyscale (centre, right), with darker shades indicating (G+C)-richer regions. SOX2 and SOX21 in Lung Epithelial Differentiation and Repair. They may also represent pseudogenes, which can be difficult in some cases to distinguish from real genes. Such genes would be hard to detect by our various techniques and would also decrease the average number of exons per gene used in the analysis above. d, Conservation near the 3 splice site. Now, the mouse is faced with "bleak December winds ensuin'" just as George, after Lennie's death, is faced with the terrible aloneness and the death of their dream with which he is left. The PubMed wordmark and PubMed logo are registered trademarks of the U.S. Department of Health and Human Services (HHS). This study aimed to investigate the susceptibility difference in AGSz and S-IRA between DBA/1 and C57BL/6 mice by profiling long noncoding RNAs (lncRNAs) and . Dyn. Consequently, Abp has been proposed to have a key role in the sexual isolation between M. musculus subspecies. We thank J. Takahashi and M. Johnston for comments on the manuscript; the Mouse Liaison Group for strategic advice; L. Gaffney, D. Leja and K.-S. Toh for graphical help; B. Graham and G. Roberts for administrative work on sequencing of individual mouse BACs; and P. Kassos and M. McMurtry for secretarial assistance. You have full access to this article via your institution. Natl Acad. Biol. In the meantime, to ensure continued support, we are displaying the site without styles A total of 7,293 amino acid variants reported to be disease-associated190 were mapped to corresponding positions in the mouse sequence. 12, 11681174 (2002), Hurst, L. D. & Smith, N. G. Do essential genes evolve slowly? 195, 477486 (1991), Tegoni, M. et al. Comparative pathway enrichment analyses between human and mouse samples reveal similarities in shared membrane trafficking and signaling pathways involved in milk fat secretion. Both measures of neutral substitution rate and SNP rate showed a significant correlation with recombination rate (Fig. Copies of LINE1 (L1) form the single largest fraction of interspersed repeat sequence in both human and mouse. Mouse seminal vesicle secretory protein of 99 amino acids (MSVSP99): characterization and hormonal and developmental regulation. & Hurst, L. D. Local similarity in evolutionary rates extends over whole chromosomes in human-rodent and mouse-rat comparisons: implications for understanding the mechanistic basis of the male mutation bias. References:A comparative encyclopedia of DNA elements in the mouse genome. 11, 15741583 (2001), Alexandersson, M., Cawley, S. & Pachter, L. SLAMcross-species GeneFinding and alignment with a generalized pair hidden Markov model. When we consider all exons rather than just coding exons, we find that 941 pairs (62%) have the same number of exons. Rev. Genotyping of additional strains reveals that the SNPs largely represent alternative alleles from M. m. domesticus and M. m. musculus, and that the blocks probably represent the distinct segmental contributions of the two subspecies to existing laboratory mouse strains. Of the approximately 5% of windows of the mammalian genome that are under selection, most do not appear to code for protein. J. 23, blue curve) using a genome-wide set of 14.3 million non-overlapping 50-bp (human) windows, each containing at least 45bp (mean 48.67bp) of aligned sequence. Over time, pseudogenes of either class tend to accumulate mutations that clearly reveal them to be inactive, such as multiple frameshifts or stop codons. J. Mol. 30, 17511756 (2002), Smith, N. G. C., Webster, M. & Ellegren, H. Deterministic mutation rate variation in the human genome. In contrast, the initial analysis of the human genome identified only three putative tRNA genes that violated the wobble rules172,173. We developed three new computer programs for dual-genome de novo gene prediction: TWINSCAN160,325, SGP2 (refs 161, 326) and SLAM162. Nature Genet. Conversely, about 78% of the predicted genes and about 81% of the exons in this catalogue were at least partially represented by TWINSCAN predictions. Accessed 5 March 2023. 281, 94100 (2001), Bain, P. A., Yoo, M., Clarke, T., Hammond, S. H. & Payne, A. H. Multiple forms of mouse 3 beta-hydroxysteroid dehydrogenase/delta 5-delta 4 isomerase and differential expression in gonads, adrenal glands, liver, and kidneys of both sexes. Second arm of research is understanding glial maturation in Autism. The mariner element is represented by elements (MMAR1 in mouse and HSMAR1 in human) that are 97% identical. Thou saw the fields laid bare an' waste, An' weary Winter comin fast, [75] An' cozie here, beneath the blast, Thou thought to dwell, Till crash! J. Mol. But no matter which organizational scheme you choose, you need not give equal time to similarities and differences. (in the press), Guig, R. et al. Nature Genet. & Wilkinson, M. F. Rapid evolution of a homeodomain: evidence for positive selection. The mouse provides a unique lens through which we can view ourselves. As a specific example of the use of the draft sequence for oncogene discovery, several groups recently used retroviral infection in mice to recover new cancer susceptibility loci. In the final stanza of To a Mouse the speaker states that the mouse is blest, compard wi him. Paired-end reads from libraries with different insert sizes were produced as previously described1 using 384-well trays to ensure linkages. And this creates a concrete argument for using comparison-oriented charts and graphs, such as Matrix and Radar Graphs. More generally, they acquire a larger ratio of non-synonymous to synonymous substitutions (KA/KS ratio; see section on proteins below) than functional genes. There is a strong positive correlation in local (G+C) content between orthologous regions in the mouse and human genomes (Fig. Sci. Trends Genet. Distinguishing regulatory DNA from neutral sites. Then when he looks forward in time he canna see or cannot see, the fears which may come for him. 19 and Table 12). The use of SNPs would allow the generation of an even denser map, which would allow mouse geneticists to fully exploit the recombinational resolution that can be achieved in large crosses. Proc. The major satellite was found in about 3.6% of the reads; this is also lower than previous estimates based on density gradient experiments, which found that major satellites comprise about 5.5% of the mouse genome, or approximately 8Mb per chromosome65. The speaker understands why this is the case and sympathizes. a, The number of lineage-specific L1 copies per megabase declines 13- to 20-fold from lowest to highest (G+C) content. What is a Research Survey? The tool has many templates to ensure a wider selection of charts. USA 85, 26532657 (1988), Sueoka, N. On the genetic basis of variation and heterogeneity of DNA base composition. This is most readily accomplished through BAC transgenesis. For you to conduct a comparative analysis, you need different types of comparison charts and graphs. It remains an important challenge to unravel the mechanistic basis and evolutionary consequences of such variation. a, Estimates are made from the REV model using all aligned sites of the given type in the chromosome. Genome 12, 590594 (2001), Purmann, L., Plass, C., Gruneberg, M., Winking, H. & Traut, W. A long-range repeat cluster in chromosome 1 of the house mouse, Mus musculus, and its relation to a germline homogeneously staining region. Epub 2022 Oct 21. 18, 337340 (2002), Castresana, J. These two classes contain relatively few exons (average 3), and thus comprise only about 12,000 exons of the 213,562 in the mouse gene catalogue. Evol. Gaining audience insights can be costly with the wrong tool. For these and other reasons, the Human Genome Project (HGP) recognized from its outset that the sequencing of the human genome needed to be followed as rapidly as possible by the sequencing of the mouse genome. The homologous genes may have been deleted in the human genome for these few cases, or they could represent the creation of new lineage-specific genes in the rodent lineagethis seems unlikely, because they show protein similarity to genes in other organisms. The key objective of this comparative chart is to help you visually depict data side by side, allowing you to see how data points stack up against one another. The real explosion, however, came with the development of recombinant DNA technology and the advent of DNA-sequence-based polymorphisms. & Nielsen, R. Estimating synonymous and nonsynonymous substitution rates under realistic evolutionary models. Copyright 1998, Kerry Walk, for the Writing Center at Harvard University, The Writing Center | Barker Center, Ground Floor. In addition, some bases outside these windows are likely to be under selection. 23 for the 50-bp windows in ancestral repeats, representing neutrally evolving DNA. These three strands of evidence are reconciled into a single gene catalogue by using heuristics to merge overlapping predictions, detect pseudogenes and discard misassemblies. Indeed, chromosome X is slightly smaller in human. The true concordance of gene structure between the two species is probably higher, because differences will be exaggerated by differential representation of alternative splice forms between the two data sets, difficulties in mapping the cDNA sequences back to the genome, and the absence of true 5 and 3 ends. 10). Morse, H. C.) 121 (Academic, New York, 1978), Haldane, J. In addition, conserved sequences probably encode non-protein-coding RNAs (which remain difficult to discern) and chromosomal structural elements. Heading independent team (7 members) exploring cell-type specificity in proteomic dysregulation seen in rat models of neurological disorders. In the final lines, he relates the mouses predicament to that experienced by all of humankind. 2014 Nov 21;346(6212):1007-12. doi: 10.1126/science.1246426. Confidence intervals were computed on the basis of the number of ancestral repeat and fourfold degenerate sites aligning in each window; points where the confidence interval does not overlap the genome-wide estimate indicate windows with significant differences in evolutionary rate. These features can sometimes be used to recognize pseudogenes, although relatively recent pseudogenes may escape such filters. By the 1700s, mouse fanciers in Japan and China had domesticated many varieties as pets, and Europeans subsequently imported favourites and bred them to local mice (thereby creating progenitors of modern laboratory mice as hybrids among M. m. domesticus, M. m. musculus and other subspecies). Some of the above differences in the nature of interspersed repeats in human and mouse could reflect systematic factors in mouse and human biology, whereas others may represent random fluctuations. Moreover, an estimated 20% of the mouse olfactory receptor homologues194 and a higher percentage of human homologues195,196 are pseudogenes, indicating that there is a dynamic interplay between gene birth and gene death in the recent evolution of this family. Anal. c, d, Interspersed repeats grouped into bins of approximately equal time periods after adjusting for the different rates of substitution in the two genomes. 9, 747750 (1999), Goodstadt, L. & Ponting, C. P. Sequence variation and disease in the wake of the draft human genome. All of the work has gone to waste as the wind has turnd the mouse out of its home. Federal and central banks worldwide use comparison charts to closely follow the global economys performance. Biol. As a starting point, let us assume that the genome size of the last common ancestor was about 2.9Gb (similar to the modern genomes of human and most other mammals) and let us focus only on large-scale insertions and deletions, ignoring nucleotide-level indels within aligned regions and lineage-specific duplications. They then search for potential exonic features, modifying the probability scores for the features according to the presence and quality of these human alignments. Curr. There are, however, several other possible reasons why this small set of mouse genes lack a human homologue. Few studies exist comparing normal cardiovascular development in mice vs. humans. Figure 14 shows this for the Zfhx1b locus, and also shows coincidence of exclusion of interspersed repeats with high conservation between human and mouse. To broaden the scope of our comparative study of mouse and human placentae across gestation beyond a handful of markers, we performed genome-wide microarray-based RNA profiling and compared gene expression both across time and between species, using 54 normal human placenta samples collected between 4 and 39 weeks gestational age, and 54 mouse The hitch-hiking effect of a favourable gene. This is an upper bound of sensitivity as some RIKEN cDNAs are probably less than full length and many tissues remain to be sampled. Lennie and George's plans are similar to that of the mouse in Robert Burns's poem. Curley's wife comes in (this can't be good). To test the accuracy of the ultracontig lengths, we compared the actual length of 675 finished mouse BAC sequences (from the B6 strain) with the corresponding estimated length from the draft genome sequence. Organizational Scheme. Rev. The B4 family resembles a fusion between B1 and ID119,120. 46, 202214 (1998), Coffin, J. M., Hughes, S. H. & Varmus, H. E. (eds) Retroviruses (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 1997), Smit, A. F. Identification of a new, abundant superfamily of mammalian LTR- transposons. An interesting case is the mariner element, which seems to have infiltrated independently both the rodent and human genomes. It should be noted that the roughly twofold higher substitution rate in mouse represents an average rate since the time of divergence, including an initial period when the two lineages had comparable rates. The single most prevalent feature of mammalian genomes is their repetitive sequences, most of which are interspersed repeats representing fossils of transposable elements. Symp. For instance, in a paper asking how the "discourse of domesticity" has been used in the abortion debate, the grounds for comparison are obvious; the issue has two conflicting sides, pro-choice and pro-life. The gene expression profile might be potentially related to this difference. Sci. 29). Genetic mapping in the mouse began with Haldane's report31 in 1915 of linkage between the pink-eye dilution and albino loci on the linkage group that was eventually assigned to mouse chromosome 7, just 2 years after the first report of genetic linkage in Drosophila. More so, you can make comparisons between categories using a highly contrasting color scheme. 2020;136:429-454. doi: 10.1016/bs.ctdb.2019.11.012. Surrounded by hard times, racial conflict, and limited opportunities, Julian, Copyright 2023 The President and Fellows of Harvard College, Writing Advice: The Barker Underground Blog, Brief Guides to Writing in the Disciplines, Writing Advice: The Harvard Writing Tutor Blog, Videos from the 2022 Three Minute Thesis Competition. When one steals one daimen-icker from a thrave or bundle of twenty-four, it is only a sma or small thing. Biophys. In total, we replaced 3,528 draft sequence contigs with 48.2Mb of finished sequence from 210 finished BACs available at the time of the assembly. MHC genotype is also known from ethological studies to influence mate selection, although the molecular mechanisms underlying this effect remain unknown. Such gene family changes represent an insight into aspects of physiology that have emerged since the last common ancestor. A. et al. This cluster, on chromosome 2, contains seminal vesicle secretory proteins that are rapidly evolving, androgen-regulated proteins involved in the formation of the copulatory plug and influence the survival and efficacy of spermatozoa209,210,211. Nature Genet. Essentially, if youre unsatisfied with the tool within a week, you can opt-out as easily as signing up for a trial. Genome Res. 374, 5356 (1995), Simon, A. M., Veyssiere, G. & Jean, C. Structure and sequence of a mouse gene encoding an androgen-regulated protein: a new member of the seminal vesicle secretory protein family. A mutation in OTOF, encoding otoferlin, a FER-1-like protein, causes DFNB9, a nonsyndromic form of deafness. 29, 201205 (2001), Van Etten, W. J. et al. & Firestein, S. The olfactory receptor gene superfamily of the mouse. With these and other loci, Haldane's original two-marker linkage group on chromosome 7 had now swelled to about 2,250 loci. We next considered how the molecular functions of domains affect their evolution. (in the press), Elnitski, L. et al. It may now be in ruins, but the speaker still wants to share what the tiny creature built. Biophys. The mouse genome sequence also has powerful applications to the molecular characterization of the somatic mutations that result in neoplasia. Cell 87, 917927 (1996), Hughes, J. F. & Coffin, J. M. Evidence for genomic rearrangements mediated by human endogenous retroviruses during primate evolution. b, Similarly, the density of CpG islands is relatively homogenous for all mouse chromosomes and more variable in human, with the same exceptions. Epub 2007 Oct 31. The mouse/human ratio has a mean at 0.91 for autosomes, but varies widely, with the mouse interval being larger than the human in 38% of cases (Fig. A. What explains the correlation among these many measures of genome divergence? 183, 494500 (1989), Davisson, M. T. & Roderick, T. H. Genetic Variants and Strains of the Laboratory Mouse (eds Lyon, M. F. & Searle, A. G.) 416427 (Oxford Univ. 167, 515 (1999), Ning, Z., Cox, A. J. J. Hum. A comparative methylome analysis reveals conservation and divergence of dna methylation patterns and functions in vertebrates Yes, because we interpret visual data faster than text and figures. This student essay consists of approximately 2pages of analysis of Of Mice and Men and To a Mouse. Placenta 23, 319 (2002), Deussing, J. et al. 12). 4, 406425 (1987), Sokal, R. & Rohlf, F. Biometry: The Principles and Practice of Statistics in Biological Research (Freeman, New York, 1995), MATH Mol. Sci. Genome 9, 491495 (1998), Ferretti, V., Nadeau, J. H. & Sankoff, D. Combinatorial Pattern Matching, 7th Annual Symposium (eds Hirschberg, D. & Myers, G.) 159167 (Springer, Berlin, 1996), Bourque, G. & Pevzner, P. A. Genome-scale evolution: reconstructing gene orders in the ancestral species. Endogenous retroviruses fall into three classes (IIII), which show a markedly dissimilar evolutionary history in human and mouse (see Fig. Nature 409, 610614 (2001), Murphy, W. J. et al. The red bar shows the location of the interferon--activated sequence-like element (GLE), which is bound by transcription factors from the STAT5a and STAT5b protein family to control expression of this gene244,245. Investigating the differences and similarities in your data is one of the most straightforward analyses you can ever conduct. Such regions comprised only a tiny fraction (<0.0001) of the total assembly, of which only half had been anchored to a chromosome. A comparative analysis between the top human (n=666) and mouse (n=873) skin-associated genes (SAGs) revealed a total of only 30.2% identity between the two lists. J. Mol. Mousehuman sequence comparisons allow an estimate of the rate of protein evolution in mammals. Although both mouse and human have discoid placentae200,201, they differ in the number and types of cell layers between the maternal and fetal blood. Dites a votre partenaire comment vous vous comparez avec vos amis et les membres de votre famille. 10, 950958 (2000), Ogata, H., Fujibuchi, W. & Kanehisa, M. The size differences among mammalian introns are due to the accumulation of small deletions. 5, 133135 (1915), Botstein, D., White, R. L., Skolnick, M. & Davis, R. W. Construction of a genetic linkage map in man using restriction fragment length polymorphisms. The placenta and the prolactin family of hormones: regulation of the physiology of pregnancy. The distribution of genes in the human genome. How to conduct comparative analysis using our easy-to-follow steps? The highly differentiated X and Y chromosomes perform a precise and specific meiotic program that includes pairing and segregation, but lacks the usual mechanisms of synapsis, recombination and chiasma formation that occur in the autosomes and also in the sex chromosomes of . 11, 8797 (2001), Shiraishi, T. et al. e, The average number of genes per window is plotted against the (G+C) content of the window for both genomes, showing that the gene density in mouse reaches the same level as in human but at a lower level of (G+C) content. Genomics 79, 225240 (2002), Afonso, S., Tovar, C., Romagnano, L. & Babiarz, B. The boss is angry that Lennie and George have shown up a day late and suspects George of taking advantage of Lennie. In human, there is evidence for at most a few active elements (HERVK10 and HERBK113 (ref. The red horizontal line represents the median and the box indicates the middle 67% of the data between the 16th and 83rd percentiles. In the human genome, the four homeobox clusters (HOXA, HOXB, HOXC and HOXD) are by far the most repeat-poor regions of the human genome, with repeat content in the range of 1%. Please continue to help us support the fight against dementia with Alzheimer's Research Charity. More so, you can efficiently conduct this analysis to investigate data points with noticeable differences and commonalities. Comparative proteomics uncovered a profibrotic and inflammatory phenotype in human and mouse obstructed kidneys . Because the latter was produced from strain 129 and other mouse strains, it is expected to differ slightly at the nucleotide level but should otherwise show good agreement. Topologically associating domains are stable units of replication-timing regulation. The mouse B1 and human Alu SINEs are unique among known SINEs in being derived from 7SL RNA; they probably have a common origin117. Why not pears and bananas? This would imply no net change in genome size in the human lineage despite the accumulation of about 700Mb of lineage-specific repeat sequence since the common ancestor (see section on repeats). Proc. This would require approximately 700Mb of deletions, implying that about 24% (700 out of 2,900) of the ancestral genome was deleted and about 76% retained in the human lineage. Frame of Reference. Internet Explorer). Previous studies have documented rapid evolution for a number of these clusters, including eosinophil-associated ribonucleases224, MHC class I227, class Cyp2d cytochromes P450 (ref. Microbiol., Washington DC, 1995), Crick, F. H. Codonanticodon pairing: the wobble hypothesis. You have to understand what issue needs to be researched and the variables that impact it. The RFX5 case is interesting, because disruption of the known mouse homologue alone does not reproduce the human disease, but may do so in conjunction with disruption of the newly identified paralogue158. Duplication boundary and evolution. Grounds for Comparison. (Domains are compact structures serving as evolutionarily conserved functional building blocks that are often assembled in various arrangements (architectures) in different proteins174.) However, proteins with KA/KS < 1 may still contain sites under positive selection, but the contribution of those sites to the KA/KS for the whole protein is offset by purifying selection at other sites185. In this respect, the mouse is unsurpassed as a model system for probing mammalian biology and human disease15,16. Proc. Sign up for the Nature Briefing newsletter what matters in science, free to your inbox daily. Again, the outliers show a clear tendency to be repeat-poor in human (see Supplementary Information). These latter cases probably represent genes that have descended from the same common ancestral gene, termed here 1:1 orthologues. Methyl-CpG is mutated by deamination to TpG, leading to approximately fivefold under-representation of CpG across the human1,95 and mouse genomes. 275, 3331433320 (2000), Peters, J. Nonspecific esterases of Mus musculus. Let's say you're writing a paper on global food distribution, and you've chosen to compare apples and oranges. In general, SSRs in which one strand is a polypurine tract and the other a polypyrimidine tract are much more common and extended in mouse than human. The third repeat class is LTR elements. Trends Genet. Slightly fewer than 2 million such sites were studied, defined in the human genome from about 9,600 human RefSeq cDNAs and aligned to their mouse orthologues. And, with his misfortune in killing Curley's wife, he is doomed to be destroyed and, with him, so is the "nest" of the dream of a ranch that he and George have--"Thy wee-bit housie, too, in ruin." A very dark and foreboding prospect. We found the location of 8,322 high-quality, coding-region SNPs from HGVbase192 within human genes using the tBLASTn computer program178 and, in turn, within the corresponding positions in mouse orthologues. What properties of chromosomal DNA could account for the variation in substitution rate? The computational pipeline produces predicted transcripts, which may represent fragmentary products or alternative products of a gene. 26, 225228 (2000), Loots, G. G., Ovcharenko, I., Pachter, L., Dubchak, I. 12). The nature and extent of conservation of synteny differs substantially among chromosomes (Fig. Hao H, Shi B, Zhang J, Dai A, Li W, Chen H, Ji W, Gong C, Zhang C, Li J, Chen L, Yao B, Hu P, Yang H, Brosius J, Lai S, Shi Q, Deng C. Mol Biomed. Biophys. 30). With knowledge of both genomes, biomedical studies of human genes can be complemented by experimental manipulations of corresponding mouse genes to accelerate functional understanding. The development of improved random mutagenesis protocols led to the establishment of large-scale screens to identify interesting new mutants, increasing the need for more rapid positional cloning strategies. Branches with significant nodes (bootstrapping value >0.7) are in black, with the remainder in blue. TWINSCAN predicted an extra 4,558 (3%) new exons not predicted by the evidence-based methods. 24). The speaker exclaims over this fact. Numerous potentially functional but non-genic conserved sequences on human chromosome 21. . Mouse regulatory DNA landscapes reveal global principles of cis-regulatory evolution. Comparative analysis is a way to look at two or more similar things to see how they are different and what they have in common. No te quites los zapatos! b, Conservation near translation start site using the same data set as in a. Although small, single-exon genes may add further to the count, the total seems unlikely to greatly exceed 30,000. The second step of filtering de novo gene predictions (by requiring the presence of adjacent exons in both species) turns out to greatly increase prediction specificity. Genomics 13, 10951107 (1992), Gardiner-Garden, M. & Frommer, M. CpG islands in vertebrate genomes. We briefly discuss RNA genes at the end of the section. Recent Prog. b, Box plot of KA/KS values for different locally duplicated, paralogous mouse-specific gene clusters. USA 99, 44714476 (2002), Paigen, K. & Eppig, J. T. A mouse phenome project. The mouse compares to Curley's wife, Crooks, Curley and Candy in that it's inevitable it will die without it's nest to protect it from the weather, as Curley's wife has already died, Crooks knows he will never realise his dream of being accepted, Curley can't live his dream of being a "real man" without a pretty wife on his arm and Candy is also facing the inevitable of having no home to go to when he loses his job. We examined the rate of deletion in the mouse genome, as measured by the fraction of non-aligning ancestral human DNA (NAanc). The neutral substitution rate has been roughly half a nucleotide substitution per site since the divergence of the species, with about twice as many of these substitutions having occurred in the mouse compared with the human lineage. Initial sequencing and comparative analysis of the mouse genome. Bootstrap values are shown at the branches. In the present research, an analysis was carried out to study the two input pointing devices, namely touchpad and mouse on the basis of throughput and location of the laptop computer. Biochemistry 28, 47794784 (1989), Miao, Y. J., Subramaniam, N. & Carlson, D. M. cDNA cloning and characterization of rat salivary glycoproteins.