黑羽番鸭CAPN1基因的克隆及生物信息学分析  

Cloning and Bioinformatics Analysis of CAPN1 Gene in Black Muscovy Duck

作　　者：李浩[1] 胡志刚[1] 陈强 张慧林[1] 刘小林[1] LI Hao;HU Zhigang;CHEN Qiang;ZHANG Huilin;LIU Xiaolin(College of Animal Science and Technology,Northwest A&F University,Shaanxi Yangling 712100,China;Anda Muscovy Duck Breeding Farm,Shaanxi Tongguan 714300,China)

机构地区：[1]西北农林科技大学动物科技学院,陕西杨陵712100 [2]陕西省潼关安大番鸭育种场,陕西潼关714300

出　　处：《中国畜牧杂志》2020年第8期177-183,共7页Chinese Journal of Animal Science

基　　金：国家水禽产业技术体系基金资助项目(CARS-42-2)。

摘　　要：本实验旨在对黑羽番鸭CAPN1基因进行克隆和生物信息学分析,以期为改善番鸭肌肉嫩度提供参考。选取黑羽番鸭为研究对象,采集腿肌组织,提取RNA,反转录为cDNA,通过同源重组方法克隆CAPN1基因的编码区序列并进行生物信息学分析。结果显示:黑羽番鸭CAPN1基因编码区序列长度为2 148 bp,编码715个氨基酸;与鸭、鸡、雉鸡、人、猪、牛、绵羊、马的序列进行同源性比较,显示核苷酸序列的同源性分别为98.70%、87.29%、86.11%、81.89%、81.62%、81.18%、81.04%、81.27%,氨基酸序列的同源性分别为99.30%、91.47%、90.63%、83.22%、83.64%、83.10%、82.96%、83.36%。系统进化分析结果与传统分类学结果一致。理化性质分析表明CAPN1蛋白相对分子质量为81 368.72 ku,理论等电点为5.82,亮氨酸使用频率最高,组氨酸使用频率最低,属于稳定性、亲水性蛋白;存在2个跨膜螺旋区,无信号肽剪切位点,主要分布于细胞质中。修饰结构分析表明,该蛋白存在59个潜在的磷酸化位点,9个O-糖基化位点和3个N-糖基化位点;二级结构中α-螺旋占比最高,β-转角占比最低;包含3个结构域;最后对CAPN1蛋白的三级结构进行了建模,模型可信度较高。The purpose of the experiment is to clone and bioinformatics analyze the CAPN1 of Muscovy duck, and to provide reference for improving the tenderness of Muscovy duck. The experiment selected black Muscovy duck as the research object, collected leg muscle tissue, extracted RNA, transcribed it into cDNA, cloned the coding region sequence of CAPN1 Gene by homologous recombination method and analyzed bioinformatics. The results showed that the length of the coding region of CAPN1 Gene was 2 148 bp, and encoded by 715 amino acids.Compared with that of duck, chicken, pheasant, human, pig, cattle, sheep and horse, the homology of nucleotide sequence was 98.70%, 87.29%, 86.11%, 81.89%, 81.62%, 81.18%, 81.04% and 81.27%, respectively;the homology of amino acid sequence was 99.30%, 91.47%, 90.63%, 83.22%, 83.64%, 83.10%, 82.96% and 83.36%, respectively. The results of phyloGenetic analysis were consistent with those of traditional taxonomy. The analysis of physical and chemical properties showed that the relative molecular weight of CAPN1 protein was 81 368.72 Da.The theoretical isoelectric point was 5.82. The frequency of leucine use was the highest, while the frequency of histidine use was the lowest. It belonged to stable and hydrophilic protein. In addition, there were two transmembrane helix regions and no signal peptide cleavage site. And, the protein is mainly distributed in the cytoplasm. The modified structure analysis showed that there were 59 potential phosphorylation sites, 9 O-glycosylation sites and 3 N-glycosylation sites in the protein. In the secondary structure, the ratio of α-helix was the highest, and the ratio of β-angle was the lowest, including 3 domains. Finally, the tertiary structure of CAPN1 protein was established with high reliability.

关 键 词：黑羽番鸭 CAPN1基因 克隆 生物信息学分析 

分 类 号：S834.2[农业科学—畜牧学]