甘肃马鹿HIF-1α基因克隆及其表达分析  

Cloning and differential expression analysis of HIF-1α gene of Gansu red deer

作　　者：张振祥 张晶晶 陈霞霞 张姗姗 田彪 陈艳霞 ZHANG Zhenxiang;ZHANG Jingjing;CHEN Xiaxia;ZHANG Shanshan;TIAN Biao;CHEN Yanxia(Gansu Engineering Laboratorv of Applied Mycology,Hexi Lniversity,Zhangye 734000,China;Qilian Mountain Ecological Research Institute,Hexi University,Zhangye 734000,China;College of Life Science and Engineering,Hexi University,Zhangye 734000,China)

机构地区：[1]河西学院甘肃省应用真菌工程实验室,甘肃张掖734000 [2]河西学院祁连山生态研究院,甘肃张掖734000 [3]河西学院生命科学与工程学院,甘肃张掖734000

出　　处：《黑龙江畜牧兽医》2021年第3期133-139,共7页Heilongjiang Animal Science And veterinary Medicine

基　　金：河西学院科研创新与应用校长基金项目(XZ2018008)。

摘　　要：为了克隆和分析甘肃马鹿缺氧诱导因子-1α(HIF-1α)基因编码区(CDS)保守区域序列,并检测其不同生长时期的差异表达,探索HIF-1α基因参与鹿茸快速生长的潜在分子机制,试验分别在花盘脱落后30天(新茸初生期)、60天(快速生长期)和90天(骨化期)时采集甘肃马鹿鹿茸,分离其顶端软骨组织,对HIF-1α基因进行克隆并对测序结果进行生物信息学分析,应用实时荧光定量PCR技术检测HIF-1α基因在不同生长时期的甘肃马鹿鹿茸软骨组织中的表达情况。结果表明:克隆所得甘肃马鹿鹿茸HIF-1α基因CDS保守区域序列长度为1 799 bp,编码599个氨基酸,与牛科动物亲缘关系较近,在物种间保守性较高。HIF-1α基因在60天甘肃马鹿鹿茸软骨组织中表达最高,显著高于30天和90天(P<0.05)。预测的miRNAs中,miR-135a的表达与HIF-1α表达趋势相反, miR-18b与HIF-1α表达趋势相同,其他3个miRNAs在生长30天和90天鹿茸软骨中的表达量与60天相比差异不显著(P>0.05)。说明试验成功克隆了甘肃马鹿HIF-1α基因CDS保守区域序列,该基因可能在miR-135a的调控下参与促进快速生长期鹿茸软骨组织的生长。In order to clone and analyze the conservative regional sequence of the CDS of HIF-1α gene in Gansu red deer, to detect the differential expression of HIF-1α at different growth periods, and to explore the potential molecular mechanism of HIF-1α gene involved in the rapid growth of deer antlers, this study collected antlers of Gansu red deer at 30 d(early stage of new antlers), 60 d(fast growth stage), 90 d(ossification period), and isolated the cartilage tissue, cloned and bioinformatics analyzed the sequence of HIF-1α. The expression of HIF-1α in the antler cartilage tissue of Gansu red deer in different growth periods was detected by qPCR technology. The results showed that HIF-1α was highly conserved among species, the sequence length of the conservative part of HIF-1α CDS region was 1 799 bp, encoded 599 amino acids, and HIF-1α in Gansu red deer was phylogenetically closer with the Bovidae. The qPCR results showed that HIF-1α had higher expression level in antler cartilage at 60 d than in 30 d and 90 d ossification periods(P<0.05). Among the predicted miRNAs, the expression of miR-135 a was negatively correlated with HIF-1α, but miR-18 b was positively correlated with HIF-1α. The expression of the other three miRNAs was not significantly different in 30 d vs. 60 d and 60 d vs. 90 d(P>0.05). This study successfully cloned the CDS of HIF-1α gene in Gansu red deer. HIF-1α gene may be involved in promoting the growth of velvet cartilage tissue in the rapid growth period under the control of miR-135 a.

关 键 词：甘肃马鹿 鹿茸 快速生长 软骨发育 HIF-1Α MIRNAS 

分 类 号：S825.2[农业科学—畜牧学] Q786[农业科学—畜牧兽医]