IFITM1基因克隆及其在不同生长阶段牦牛各组织中的表达分析  

Cloning of IFITM1 gene and analysis of its expression in yak tissues at different growth stages

作　　者：王海鹏 李娟[1] 王利[1] 郑姚 张玲[1] WANG Hai-peng;LI Juan;WANG Li;ZHENG Yao;ZHANG Ling(Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization,Southwest Minzu University,Ministry of Education and Sichuan Province,Chengdu 610041,China)

机构地区：[1]西南民族大学青藏高原动物遗传资源保护与利用教育部和四川省重点实验室,四川成都610041

出　　处：《中国预防兽医学报》2021年第2期139-144,190,共7页Chinese Journal of Preventive Veterinary Medicine

基　　金：西南民族大学研究生创新型项目(CX2020SZ30);四川省留学人员科技活动择优资助项目(2019)。

摘　　要：为鉴定牦牛IFITM1基因并分析其在不同生长阶段牦牛5种组织中的转录水平和蛋白表达量。本研究采用根据普通牛IFITM1基因设计的引物,PCR扩增牦牛IFITM1基因并利用生物信息学软件分析其序列,结果显示,获得牦牛546 bp的IFITM1基因cDNA序列,其中编码(CDS)区为375 bp,编码124个氨基酸残基。牦牛IFITM1基因序列与普通牛的同源性最高,为98.7%。进一步采用荧光定量PCR检测IFITM1基因在不同生长阶段牦牛各组织中的转录水平,结果显示,IFITM1在1日龄、15月龄和5岁龄牦牛的心脏、肝脏、脾脏、肺脏和肾脏中均有不同程度的转录水平,在肝脏中的转录水平均显著高于在脾脏、肺脏和肾脏中的转录水平(p<0.05),随着年龄的增长IFITM基因在牦牛肝脏中的转录水平呈现先上升后下降的趋势,15月龄相对于1日龄时的转录水平显著升高(p<0.01),随着年龄的增长IFITM1基因的转录水平在耗牛肺脏中呈现持续下降的趋势,而在脾脏和肾脏中呈现先下降后稳定的趋势,且5岁龄相对于1日龄时的转录水平均显著降低(p<0.01)。同时采用免疫组化技术检测IFITM1蛋白在牦牛肝脏中的定位及表达。结果显示,IFITM1蛋白在3个生长阶段牦牛的肝脏中均有表达并定位于细胞膜,且该蛋白表达量随着年龄的增长呈现先下降后上升的趋势。本研究为深入探究IFITM1蛋白的功能奠定基础。To identify and analyze the yak IFITM1 gene transcription and protein expression levels in five yak tissues at different growth stages,primers were designed based on the common cattle IFITM1 gene.Subsequently,the yak IFITM1 gene was amplified by PCR and the sequence was analyzed using bioinformatics software.The results showed that the cDNA sequence of the yak IFITM1gene was 546bp,of which the coding(CDS)region was 375bp,encoding 124 amino acid residues.The nucleotide sequenc of IFITM1 of yak shares the highest homology to cattle,which is 98.7%.Further,quantitative real-time RT-qPCR was used to detect the transcription level of the IFITM1 gene in various yak tissues at different growth stages.The results showed that the IFITM1 has different transcription levels in the heart,liver,spleen,lung and kidney tissues of one-day-old,fifteen-month-old and five-year-old yak.The transcription level in liver tissue is significantly higher than that in spleen,lung and kidney tissues(p<0.05).With the increase of age,the transcription level of IFITM1 in liver tissue increased at first and then decreased.The transcriptional level of IFITM1 at fifteen-month-old was significantly higher than that at one day old(p<0.01).With the increase of age,the transcription level decreased continuously in lung tissue,decreased first and then stabilized in spleen and kidney tissue,and significantly reduced in five-year-old compared with one-day-old(p<0.01).Simultaneously,an immunohistochemical technique was used to detect the localization and expression of IFITM1 protein in the yak liver.IFITM1 protein is expressed in the yak liver in three growth stages and is located in the cell membrane,and the expression of this protein shows a trend of first decreasing and then increasing with age.This study laid the foundation for the in-depth research of IFITM1 protein function.

关 键 词：牦牛 IFITM1 基因克隆 组织表达 免疫组化 

分 类 号：S852.4[农业科学—基础兽医学]