西藏小型猪MSTN基因分子克隆及序列分析
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广东省科技计划项目(2012B011000004、2012B010300001);广东省科技厅科技基础条件建设项目(2010B060500001);国家科技部国际合作项目(2011DFA33290)。


Cloning and sequence analysis of Tibet mini-pig's MSTN gene
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    摘要:

    目的 扩增西藏小型猪的肌肉生长抑制素(MSTN)基因编码区全长序列,并对序列进行生物信息学分析。方法 提取西藏小型猪肝脏组织的RNA,反转录成为cDNA,利用RT-PCR方法扩增MSTN基因编码区全长序列,将纯化的片段与pMD19-T载体连接,转化大肠杆菌DH5α,筛选阳性克隆并测定其序列,利用生物信息学方法分析其序列特征,并与其他12种物种构建系统进化树。结果 西藏小型猪MSTN基因编码区全长1128 bp,编码375个氨基酸,氨基酸序列同源性分析表明,与版纳微型猪MSTN氨基酸序列相似性最高,为99%。结论 MSTN基因西藏小型猪除了与鸡和斑马鱼亲缘关系较远外,与人、犬、版纳微型猪、绵羊、山羊、牛、马、黑猩猩、大鼠、小鼠的关系都较为接近,其中与版纳微型猪的亲缘关系最为接近。

    Abstract:

    Objective To amplify the encoding full-length sequence of Tibet mini-pigs myostatin (MSTN) gene and analysis the sequence by bioinformatics software. Method The RNA of liver tissues from Tibet mini-pig was extracted, and reversely transcribed into cDNA. The gene coding region sequence of myostatin gene was amplified through RT-PCR, and then the purified product of PCR was ligated with a pMD19-T and transferred into the bacterium DH5α for replication. The positive clones were screened and sequenced. The sequence characters were analyzed by using bioinformatics method and phylogeny evolution tree was constructed with other twelve species. Results The coding redion of MSTN gene was 1128bp, and coded 375 amino acids. The amino acid homology analysis showed that the homology rate of amino acid sequence was 99%. Conclusions Molecular phylogeny evolution indicated that it had a close relation with human, dog, banna minipig, sheep, goat, cattle, horse, chimpanzee, rat, mouse except chick and zebrafish, and the most closely related with banna minipig.

    参考文献
    [1] Mcpherron A C,Lee S J. Double muscling in cattle due to mutations in the Myostatin gene[J]. Proc Natl Acad Sci USA,1997,94(23):12457-12461.
    [2] 岳敏,顾冬生,李洪涛,等. 西藏小型猪MSTN基因5'调控区的酶切多态性分析[J]. 实验动物与比较医学,2008,28(3):155-159.
    [3] 王伟,连林生,李继中,等.猪MSTN基因生物信息学分析[J]. 安徽农业科学,2012,40(10):5943-5945.
    [4] 刘晓琴,马喜山,唐中林,等. 猪MSTN基因的多态性和生长性状关联分析[J]. 畜牧兽医学报,2013,44(7):1063-1069.
    [5] Holmes J H G,Ashmore C P.A histochemical study of mutable site fiber type and size in normal and double muscled cattle[J]. Growth,1972,36: 351- 352.
    [6] Kollias H D,McDermott C.Transforming growth factor-β and myostatin signaling in skeletal muscle[J]. J Appl Physiol,2008,104:579- 587.
    [7] Hill J J,Davies M V,Pearson A A, et al. The myostatin propeptide and the follistatin-related gene are inhibitory binding proteins of myostatin in normal serum[J]. J Biol Chem,2002,277(43) :40735 - 40741.
    [8] Ji S,Losinski R L,Cornelius S G, et al. Myostatin expression in porcine tissues: tissue specificity and developmental and postnatal regulation[J]. American Journal of Physiology,1998,275:1265- 1273.
    [9] 乔宪凤,刘西梅,华文君,等. 猪肌肉生成抑制素(MSTN)基因cDNA克隆及序列分析[J]. 江西农业学报,2010,22(10):130-132.
    [10] 李洪涛,顾为望,袁进,等. 实验用西藏小型猪原代和第一代间部分血液指标比较[J]. 郑州大学学报(医学版),2008,43(1)63-65.
    [11] 王玉珏,郭凯,陈驰,等. 不同辐射剂量对西藏小型猪小肠线粒体的影响[J]. 南方医科大学学报,2012,32(10):1445-1450.
    [12] 黄威,毛莹莹,刘薇,等. 3种腺相关病毒介导的绿色荧光蛋白对西藏小型猪胚胎成纤维细胞的转染效率[J]. 南方医科大学学报,2012,32(6):857-861.
    [13] 李洪涛,吴清洪,袁进,等. 实验用西藏小型猪线粒体DNA控制区变异类型及其血液指标比较研究[J]. 南方医科大学学报,2009,29(8):1626-1628.
    [14] M cPherron A C. et al. Regulation of skeletal muscle mass in m ice by a new TGF-B superfam ilymember[J]. N ature, 1997,(387):83- 90.
    [15] M cPherron A C, L ee S J. The transforming growth factor beta-super family [A]. In: B. D. L eRo ith and C. Bondy,edito rs. Grow th facto rs and cytok ines in health and disease.(Volume 1) [M]. JA I P ress Inc. Greenw ich, Connecticut,U SA, 1996. 357-393.
    [16] Bogdanovich S, Krag TO, Barton ER, et al. Functional improvement of dystrophic muscle by myostatin blockade[J].Nature,2002 Nov 28:420(6914):418-421.
    [17] 张利媛,岳文斌. MSTN基因的研究进展[J]. 草食家畜,2006(132):1-4.
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吴清洪,岳敏,田雨光,王玉珏,徐名衬,顾为望.西藏小型猪MSTN基因分子克隆及序列分析[J].中国比较医学杂志,2015,25(3):18~22.

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  • 最后修改日期:2015-02-27
  • 在线发布日期: 2015-04-22
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