龙胜凤鸡IGF2BP1基因的克隆、序列分析和过表达载体构建  

Bioinformatics Analysis and Construction on Overexpression Vector of IGF2BP1 in Longsheng-Feng Chicken

作　　者：李现超 杨祝良[1] 孙甜甜 杨雪琴 张佳仪 黄超 粟永春 杨秀荣[1] LI Xianchao;YANG Zhuiang;SUN Tiantian;YANG Xueqin;ZHANG Jiayi;HUANG Chao;SU Yongchun;YANG Xiurong(College of Animal Science and Technology,Guangxi University,Nanning,Guangxi 530004;Guangxi Jinling Agriculture and Animal Husbandry Group Co.,Ltd.,Nanning,Guangxi 530028)

机构地区：[1]广西大学动物科学技术学院,广西南宁530004 [2]广西金陵农牧集团有限公司,广西南宁530028

出　　处：《中国家禽》2024年第2期119-124,共6页China Poultry

基　　金：广西创新驱动发展专项(桂科AA17204027);广西重点研发计划项目(桂科AB21075010);广西科技基地和人才专项(桂科AD19245076)。

摘　　要：为研究胰岛素样生长因子2 mRNA结合蛋白1(Insulin-like growth factor 2 mRNAbinding protein 1,IGF2BP1)基因的序列和在鸡肌肉发育中的功能,研究克隆IGF2BP1基因并进行生物信息学分析,构建IGF2BP1的过表达载体pEGFP-IGF2BP1,在成肌细胞验证载体的功能。结果显示:龙胜凤鸡IGF2BP1基因CDS区序列全长1 731 bp,编码576个氨基酸,与参考序列相比,存在6处同义突变;物种间同源性分析显示,龙胜凤鸡IGF2BP1基因与原鸡(Gallus gallus)、火鸡(Meleagris gallopavo)、绿头鸭(Anas platyrhynchos)、猪(Sus scrofa)、奶牛(Bos taurus)、小鼠(Mus musculus)的相似性分别为99.7%、94.7%、92.5%、82.4%、82.8%、82.7%;蛋白质二级、三级结构主要含有无规卷曲(45.83%)、α螺旋(27.26%)和延伸链(20.83%);经过酶切和测序鉴定显示,成功构建了pEGFP-IGF2BP1质粒,并且该质粒可在成肌细胞中表达绿色荧光,qRT-PCR结果显示IGF2BP1在试验组(转染pEGFP-IGF2BP1)的表达量极显著高于对照组(转染pEGFP-N1)(P<0.01)。结果表明,成功克隆IGF2BP1基因,构建的IGF2BP1基因过表达载体在成肌细胞中成功表达。To investigate the sequence of insulin-like growth factor 2 mRNA-binding protein 1(IGF2BP1)gene and its function in chicken muscle development,in this study,the IGF2BP1 gene was cloned and bioinformatics analysis was performed.The IGF2BP1 overexpression vector pEGFP-IGF2BP1 was constructed,and its function was verified in myoblast.The results showed that the total length of the CDS region of IGF2BP1 gene in Longsheng-Feng chicken was 1731 bp,encoding 576 amino acids,and there were 6 synonymous mutations compared with the reference sequence.The homology analysis among species showed that the IGF2BP1 gene of Longsheng-Feng chicken had 99.7%,94.7%,92.5%,82.4%,82.8%and 82.7%homology with the Gallus gallus,Melagris gallopavo,Anas platyrhynchos,Sus scrofa,Bos tauru and Mus muculus,respectively.The secondary and tertiary structures of proteins mainly contain random coils(45.83%),αhelices(27.26%)and extended chains(20.83%).After enzyme digestion and sequencing identification,pEGFP-IGF2BP1 plasmid was successfully constructed,and the plasmid could express green fluorescence in myoblast.The qRT-PCR results showed that the expression of IGF2BP1 in the test group(transfected with pEGFP-IGF2BP1)was extremely significantly higher than that in the control group(transfected with pEGFP-N1)(P<0.01).The results indicated that the IGF2BP1 gene was successfully cloned and the constructed IGF2BP1 gene over expression vector was successfully expressed in myoblasts.

关 键 词：龙胜凤鸡 IGF2BP1基因 生物信息学 过表达载体 

分 类 号：S831.2[农业科学—畜牧学]