中国新吉细毛羊LHx2基因cDNA克隆与序列分析.doc

中国新吉细毛羊LHx2基因cDNA 克隆与序列分析 王沙沙，金海国2，曹阳2，鲁承1，张雪梅1 ，孙福亮1*，张立春2* （1.延边大学 农学院，吉林 延吉 133002；2.吉林省农业科学院 畜牧分院，吉林 公主岭 136100） 摘要：此研究以供试羊皮肤组织中提取的总RNA为模板，根据GenBank中登陆的绵羊LHx2基因序列设计引物，通过PCR方法扩增RT-PCR合成的LHx2基因编码区第一链cDNA，回收、纯化反应产物，将目的基因与pMD-18T载体连接、转化，获得阳性重组质粒，成功克隆新吉细毛羊LHx2基因cDNA序列，测序长为1 215 bp；基因同源性分析各物种同源性均在80%以上；该基因编码405个氨基酸残基，蛋白同源性分析各物种同源性差异较大，蛋白二级结构分析显示LHx2蛋白为疏水性蛋白、柔韧性较高，蛋白结构域分析结果表明克隆的基因较为完整。研究结果为深入研究新吉细毛羊LHx2基因提供理论基础。 关键词：新吉细毛羊、LHx2基因、克隆、序列分析 Cloning and sequence analysis of LHx2 gene cDNA from China XinJi fine wool sheep WANG Sha-sha1, JIN Hai-guo2, CAO Yang2, LU Cheng1, ZHANG Xue-mei1, SUN Fu-liang1*, ZHANG Li-chun2* (1.College of Agricultural ,Yanbian University ,Yanji,Jilin 133002,China；2.Branch of Animal Husbandry，Jilin Academy of Agricultural Science，Gongzhuling,Jilin 136100，China) Abstract: The test through for RNA extraction test sheep skin tissue as templates, primers were designed according to LHx2 gene sequence published in GenBank sheep, RT-PCR method of synthesis of LHx2 gene encoding region of the first chain cDNA using amplified by PCR, the recovery and purification of the reaction product, to get the recombinant plasmids, by putting the target gene and pMD-18T vector cloned and transformed. This experiment successfully cloned and sequenced in Xinji fine wool sheep LHx2 gene cDNA sequence, the length is 1 215 bp. Each species homology was more than 80% analysis of gene homology; the gene encoding a 405 amino acid protein, homology analysis showed that the homology of the larger species differences, the two level structure analysis predict that the protein is the hydrophobic and the protein flexibility high, domain analysis showed that the cloned gene is complete. The test provides a theoretical basis for the further study of Xinji fine wool sheep LHx2 gene. Key words：Xinji fine wool sheepLHx2 gene; Clone 新吉细毛羊是细毛羊新品种，由新疆畜牧科学院、新疆农垦科学院及吉林省农业科学院培育成功[1]，主要特点具有自身独特的被毛细度、稳定的遗传性能[2]。LHx2基因又被称为LH-2，是组成LIM同源结构域(LIM homeodomain，简称LIM-HD)转录因子家族的成员[3]。LHx2基因包含5个外显