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| 天祝白牦牛LF基因克隆及分子特征 | |
| 引用本文： | 裴杰,阎萍,姬国红,厍睿,冯瑞林,朱新书,梁春年,郭宪,曾玉峰,包鹏甲.天祝白牦牛LF基因克隆及分子特征[J].中国农业科学,2009,42(3):1030-1038. |
| 作者姓名： | 裴杰 阎萍 姬国红 厍睿 冯瑞林 朱新书 梁春年 郭宪 曾玉峰 包鹏甲 |
| 作者单位： | 1. 中国农业科学院兰州畜牧与兽药研究所,兰州,730050 2. 甘肃农业大学草业学院,兰州,730070 3. 西北民族大学生命科学与工程学院,兰州,730030 |
| 基金项目： | 中央级公益性科研院所基本科研业务费专项基金 |
| 摘 要： | 【目的】哺乳动物乳铁蛋白（Lactoferrin，LF）在抗菌、抗病毒、抗肿瘤、调节免疫等方面起着重要作用。本试验对天祝白牦牛乳铁蛋白基因的编码区进行克隆和分子特征分析。【方法】以处于干乳期的天祝白牦牛乳腺组织为材料，通过RT-PCR克隆了包含LF基因编码区的cDNA序列（GenBank收录号为EU547252）;将克隆获得的天祝白牦牛LF基因的cDNA与奶牛相应序列进行比对;对天祝白牦牛LF蛋白（GenBank收录号为ACB29795）与其它物种LF蛋白进行序列比对和进化树分析;对牦牛LF蛋白的特性和结构进行预测。【结果】克隆获得天祝白牦牛LF基因cDNA片段长度为2 344 bp，其中LF基因编码区全长2 124 bp，编码708个氨基酸;序列分析显示，克隆获得的牦牛cDNA序列与奶牛该序列存在15个碱基的变异，其中位于LF基因编码区的变异有12个，这些突变造成4个氨基酸变异;天祝白牦牛LF蛋白与奶牛、人、小鼠、山羊、绵羊、猪、狗、马、骆驼、猩猩、鸡LF蛋白氨基酸相似度分别为99.4%、69.5%、63.5%、92.4%、91.5%、72.9%、69.1%、72.6%、75.3%、69.5%和50.9%;各物种LF蛋白进化树符合物种进化规律;同源建模预测LF 3D模型显示，LF为多肽链在二级结构基础上折叠形成2个极相似的、对称的球状叶，即N叶和C叶，中间由1段对蛋白酶敏感的α-螺旋连接，呈“二枚银杏叶型”结构。【结论】从天祝白牦牛克隆获得LF基因编码区全长序列并揭示了其分子特征，为牦牛LF蛋白基因工程和蛋白质功能的研究奠定了基础。 |
| 关 键 词： | 牦牛 LF基因 克隆 分子特征 > |
| 收稿时间： | 2008-6-5 |
cDNA Cloning of lactoferin and Its Characteristics in the Tianzhu White Yak |
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| PEI Jie,YAN Ping,JI Guo-hong,SHE Rui,FENG Rui-lin,ZHU Xin-shu,LIANG Chun-nian,GUO Xian,ZENG Yu-feng,BAO Peng-jia.cDNA Cloning of lactoferin and Its Characteristics in the Tianzhu White Yak[J].Scientia Agricultura Sinica,2009,42(3):1030-1038. | |
| Authors: | PEI Jie YAN Ping JI Guo-hong SHE Rui FENG Rui-lin ZHU Xin-shu LIANG Chun-nian GUO Xian ZENG Yu-feng BAO Peng-jia |
| Institution: | Lanzhou Institute of Animal Science and Veterinary Parmaceutics, China Academy of Agricultural Sciences |
| Abstract: | 【Objective】 Lactoferrin play a very important role in antibiosis, antivirus, antitumor and immunomodulation for mammal. The coding region of LF gene of Tian Zhu white yak was cloned, and it’s molecular characteristics were analyzed in the study. 【Method】 Using mammary tissue of Tian Zhu white yak as material, the cDNA sequence containing coding region of LF gene was cloned by RT-PCR. The cDNA of LF gene of Tian Zhu white yak obtained by cloning was compared to corresponding sequence of cow. Sequence alignment and evolutionary tree were analyzed on Tian Zhu white yak and other species. The structure and properties of LF gene of yak were predicted. 【Result】 The 2 344 bp length cDNA of LF gene of Tian Zhu white yak was cloned for the first time, and the length of coding region of LF gene is 2 124 bp, coding 708 amino acids. There are 15 mutations in the cDNA between yak and cow, 12 mutations being in coding region of LF gene, causing 4 amino acid mutations in LF protein. There is high homeology of LF protein among different species. Amino acid similarities of LF protein compared yak to cow, human, mouse, goat, sheep, pig, dog, horse, camel, chimpanzee and chicken are 99.4%, 69.5%, 63.5%, 92.4%, 91.5%, 72.9%, 69.1%, 72.6%, 75.3%, 69.5% and 50.9%, respectively. The evolutionary tree of all species complies with laws of evolution. Homogenous 3D model of LF obtained by predict shows that LF protein forms two similar and symmetrical leaves, namely N-leaf and C-leaf, connected by an Alpha helix impressionable to prolease, and LF protein presents “two-folium-ginkgo-shaped” structure. 【Conclusion】 The coding region of LF gene in Tian Zhu white yak was cloned for the first time, and its molecular characteristics were shown. This work has laid an important foundation for researches of genetic engineering and protein function of LF protein of yak. |
| Keywords: | yak LF gene clone molecular characteristics |
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