cDNA Cloning, Sequence Analysis and Tissue Expression of Gene MSH4 and MSH5 in Goats

doi:10.13560/j.cnki.biotech.bull.1985.2016.03.016

Abstract

Abstract: The total RNA of ovary, endometrium, oviduct and pituitary extracted from 5 Tibetan goats and 5 Jintang black goats in the period of estrus were used to clone and analyze cDNA sequences of gene MSH4 and MSH5 by RT-PCR, and their expression levels in different tissues were determined by real-time PCR. The result showed that the coding region of gene MSH4 from Tibetan goat and Jintang black goat were 2 499 bp, encoding 832 amino acids；there were 5 base variations between two breeds, leading to 3 differences of amino acid. The coding region of gene MSH5 from Tibetan goat and Jintang black goat were 2 496 bp, encoding 831 amino acids；there were 9 base variations between two breeds, leading to 5 differences in amino acid. The nucleotide sequences of coding region of gene MSH4 from Tibetan goat were 99.8%, 99.8%, 99.4%, 98.1%, 94.4%, 85.1%, 84.7% and 93.5% homology with that of Jintang black goat, Capra hircus, Ovis aries, Bos taurus, Equus caballus, Mus musculus, Rattus norvegicus, and Homo sapiesn, respectively. The nucleotide sequences of coding region of gene MSH5 from Tibetan goat were 99.6%, 99.6%, 97.3%, 88.0%, 85.8%, 85.3% and 90.2% homology with that of Jintang black goat, C. hircus, B. taurus, Canis lupus familiaris, M. musculus, R. norvegicus, and H. sapiens, respectively. The mRNA of gene MSH4 and MSH5 were all expressed in ovary, endometrium, oviduct and pituitary of two breeds of goat, but there was no significant difference between two breeds(P>0.05). The results reveal that both gene MSH4 and MSH5 are conservative in the course of animal evolution, and the role of their effect on prolificacy of goats needs to be further studied.

Key words: Tibetan goat, Jintang black goat, MSH4, MSH5, cloning, qPCR

ZHENG Jie, LIU Shuang, LUO Bin, HU Liang, YANG Ke-wei, ZI Xiang-dong. cDNA Cloning, Sequence Analysis and Tissue Expression of Gene MSH4 and MSH5 in Goats[J]. Biotechnology Bulletin, 2016, 32(3): 98-104.

References

[1]王杰, 王永, 欧阳熙, 等. 藏山羊研究[J]. 中国畜牧杂志, 1993, 29(1)：10-13.
[2]许德贵, 张家明, 唐诗军, 等. 金堂黑山羊生产性能研究[J]. 四川畜牧兽医, 2006, 33(11)：25-26.
[3]Notter DR. Genetic improvement of reproductive efficiency of sheep and goats[J]. Anim Reprod Sci, 2012, 130(3-4)：147-151.
[4]胡亮, 龙石太, 吴宪红, 等. 川中黑山羊OPN基因cDNA克隆及生物信息学分析[J]. 江苏农业科学, 2014, 42(5)：34-36.
[5]龙石太, 胡亮, 吴宪红, 等. 川中黑山羊BMP2、BMP4基因cDNA克隆及生物信息学分析[J]. 江苏农业科学, 2014, 42(3)：19-23.
[6]Nagaoka SI, Hassold TJ, Hunt PA. Human aneuploidy：mechanisms and new insights into an age-old problem[J]. Nat Rev Genet, 2012, 13：493-504.
[7]Schroering AG, Edelbrock MA, Richards TJ, et al. The cell cycle and DNA mismatch repair[J]. Exper Cell Res, 2007, 313(2)：292-304.
[8]Jirieny J. The multifaceted mismatch-repair system[J]. Nat Rev Mol Cell Biol, 2006, 7(5)：335-346.
[9]Eisen JA. A phylogenomic study of the MutS family of proteins[J]. Nucleic Acids Res, 1998, 26(18)：4291-4300.
[10]Fishel R, Wilson T. MutS homologs in mammalian cells[J]. Curr Opin Genet Dev, 1997, 7(1)：105-113.
[11]Karen VM, Cassandra J, Yashna T, et al. Separable crossover-promoting and crossover-constraining aspects of Zip1 activity during budding yeast meiosis[J]. PLoS Genet, 2015, 11(6)：e1005335.
[12]骆骅. 黄牛和犏牛睾丸组织中减数分裂同源重组基因表达、克隆与启动子区甲基化分析[D]. 南京：南京农业大学, 2013.
[13]禄婷婷. 错配修复蛋白基因多态性与少精、无精症的相关研究[D]. 长沙：中南大学, 2009.
[14]Ramaswamy R, Narayanaswamy S, Koodali TN. Structural insights into saccharomyces cerevisiae Msh4-Msh5 complex function using homology modeling[J]. PLoS One, 2013, 8(11)：e78753.
[15]Snowden T, Acharya S, Butz C, et al. hMSH4-hMSH5 recognizes Holliday Junctions and forms a meiosis-specific sliding clamp that embraces homologous chromosomes[J]. Mol Cell, 2004, 15(3)：437-451.
[16]禄婷婷, 徐克前. 一组多功能的错配修复蛋白[J]. 生命的化学, 2008, 165(6)：696-700.
[17]Wu XL, Xu KQ, Her CT. The Role of muS homologues MSH4 and MSH5 in DNA metabolism and damage response[M]// Kusic-Tisma J. DNA replication and related cellular processes. Rijeka, Croatia：In Tech Press, 2001：87-110.
[18]Her C, Zhao N, Wu X, et al. MutS homologues hMSH4 and hMSH5：diverse functional implications in humans[J]. Front Biosci, 2007, 12(3)：905-911.
[19]Edelmann W, Cohen PE, Kneitz B, et al. Mammalian MutS homologue 5 is required for chromosome pairing in meiosis[J]. Nature Genetics, 1999, 21：123-127.
[20]Kneitz B, Cohen PE, Avdievich E, et al. MutS homolog 4 localization to meiotic chromosomes is required for chromosome pairing during meiosis in male and female mice[J]. Genes Dev, 2000, 14(9)：1085-1097.
[21]De Vries SS, Baart EB, Dekker M, et al. Mouse MutS-like protein Msh5 is required for proper chromosome synapsis in male and female meiosis[J]. Genes Dev, 1999, 13(5)：523-531.
[22]Ehrenstein MR, Neuberger MS. Deficiency in Msh2 affects the efficiency and local sequence specificity of immunoglobulin class-switch recombination：parallels with somatic hypermutation[J]. EMBO J, 1999, 18(12)：3484-3490.
[23]Shenker NS, Polidoro S, van Veldhoven K, et al. Epigenome-wide association study in the European Prospective Investigation into Cancer and Nutrition(EPIC-Turin)identifies novel genetic loci associated with smoking[J]. Hum Mol Genet, 2013, 22(5)：843-851.
[24]Kazma R, Babron MC, Gaborieau V, et al. Lung cancer and DNA repair genes：multilevel association analysis from the International Lung Cancer Consortium[J]. Carcinogenesis, 2012, 33(5)：1059-1064.
[25]Pfaffl MW. A new mathematical model for relative quantification in real-time RT-PCR[J]. Nucleic Acids Res, 2001, 29(9)：e45.
[26]胡亮, 字向东, 卢建远, 等. 多胎和单胎山羊品种Bcl-2和Bax基因的克隆及组织表达研究[J]. 畜牧与兽医, 2015, 47(1)：28-32.
[27]Bocker T, Barusevicius A, Snowden T, et al. hMSH5：a human MutS homologue that forms a novel heterodimer with hMSH4 and is expressed during spermatogenesis[J]. Cancer Res, 1999, 59(4)：816-822.
[28]Her C, Doggett NA. Cloning, structural characterization, and chromosomal localization of the humanorthologue of Saccharomyces cerevisiae MSH5 gene[J]. Genomics, 1998, 52(1)：50-61.
[29]Moens PB, Kolas NK, Tarsounas M, et al. The time course and chromosomal localization of recombination-related proteins at meiosis in the mouse are compatible with models that can resolve the early DNA-DNA interactions without reciprocal recombination[J]. J Cell Sci, 2002, 115(8)：1611-1622.
[30]Ross-Macdonald P, Roeder GS. Mutation of a meiosis-specific MutS homolog decreases crossing over but not mismatch correction[J]. Cell, 1994, 79(6)：1069-1080.
[31]Her C, Wu X, Griswold MD, et al. Human MutS homologue MSH4 physically interacts with von Hippel-Lindau tumor suppressor-binding protein 1[J]. Cancer Res, 2003, 63(4)：865-872.