| 川西高原黑唇鼠兔乳酸脱氢酶C基因cDNA的克隆及序列分析（摘要） |
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| 引用本文： | 贺庆华,徐亚欧,郑玉才,陈锋,张文磊.川西高原黑唇鼠兔乳酸脱氢酶C基因cDNA的克隆及序列分析（摘要）[J].农业科学与技术,2009,10(6):63-66. |
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| 作者姓名： | 贺庆华 徐亚欧 郑玉才 陈锋 张文磊 |
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| 作者单位： | 西南民族大学生命科学与技术学院,四川成都,610041 |
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| 基金项目： | 国家民委资助课题，四川省应用基础研究，四川省学术文化与技术领袖基金.Supported by State Ethnic Affairs Commission of P;，Applicable and Fundamental Research Funds of Sichuan Province，Academic Culture and Technol Leaders in Sichuan Province Foundation |
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| 摘 要： | 目的]为研究以鼠兔LDH-C4为靶蛋白的节育型鼠药奠定基础。方法]黑唇鼠兔属于兔形目,鼠兔属,在GenBank数据库里没有兔形目物种LDH-C基因的相关序列,故采用设计简并引物的方法进行克隆。设计的3条简并引物序列为：D-Ps：5′-atgtcmacygtcaaggagcagct-3′：D-Pa1：5′-gcagayacabtgtaatcttttcc-3′;D-Pa2：5′-ttacarccacttccratyac-3′。用反转录生成的cDNA作为模板,采用巢式PCR法克隆LDH-C基因的EST,再根据EST序列设计了2条基因特异引物,采用3′RACE技术克隆LDH-C基因的3′端,2条基因特异引物为Pika-Ps1：5-cttgcccttgttgatgttgcaga-3和Pika-Ps2：5-ggatcttcaacatggcagtct-3。核酸序列的分析、拼接和相似性搜索采用Vector NTI Suite 9软件,系统发生树的构建用Mega 4.0软件,采用邻接法。结果]黑唇鼠兔的LDH-C基因的cDNA全长1 498 bp,其中ORF长996 bp,编码332个氨基酸,3′UTR长486 bp。ORF序列比对显示LDH-C基因在大的分类单位上均很保守。系统树分析显示黑唇鼠兔的LDH-C基因和灵长类及偶蹄目的遗传距离较近,和啮齿目较远。结论]成功的克隆得到了黑唇鼠兔的LDH-C基因的cDNA序列。
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| 关 键 词： | LDH-C基因 RACE EST 序列分析 |
Cloning and Sequence Analysis of Lactate Dehydrogenase C(LDH-C)Gene from Black-lipped Pika in Western Sichuan Plateau |
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| HE Qing-hua,XU Ya-ou,ZHEN Yu-cai,CHEN Feng,ZHANG Wen-lei.Cloning and Sequence Analysis of Lactate Dehydrogenase C(LDH-C)Gene from Black-lipped Pika in Western Sichuan Plateau[J].Agricultural Science ＆ Technology,2009,10(6):63-66. |
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| Authors: | HE Qing-hua XU Ya-ou ZHEN Yu-cai CHEN Feng ZHANG Wen-lei |
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| Institution: | ( College of Life Science and Technology, Southwest University for Nationalities, Chengdu 610041 |
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| Abstract: | Objective] The aim was to lay a foundation for research of contraceptive rodenticide with LDH-C4 as target protein. Method] EST sequence of LDH-C gene from black-lipped pika was cloned by PCR with degenerate primers; then the full length open reading frame （ORF） and 3＇UTR sequence were cloned by RACE technique. Result] The full length cDNA was 1 498 bp containing an ORF of 996 bp and a 3＇UTR of 486 bp. The ORF encoded a polypeptide of 332 amino acids. The alignment of LDH-C gene ORF nucleotide sequences from different species showed that the gene was conserved even between large taxons. The phylogenic tree showed that black-lipped pika LDH-C was closer to prima- tes and artiodactyla than to rodents. Conclusion] cDNA sequence of LDH-C gene from black-lipped pika was cloned successfully. |
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| Keywords: | RACE EST LDH-C gene RACE EST Sequence analysis |
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