牦牛KLF3基因克隆及组织表达分析

Cloning and Tissue Expression Analysis of KLF3 Gene in Yak(Bos grunniens)

旨在克隆牦牛KLF3基因序列,并获得其生物学特征,同时阐明其组织表达规律。选取4-6周岁的健康麦洼牦牛6头,采集脾、肺、肾、皮下脂肪和背最长肌组织样品,提取组织总RNA,利用RT-PCR方法克隆KLF3基因序列,同时利用荧光定量PCR(q PCR)技术检测该基因在不同组织中的表达情况。结果表明,获得牦牛KLF3基因序列1 137 bp(Gen Bank登录号:KX964630),其中CDS为1 041 bp,5'UTR 32 bp和3'UTR 64 bp,编码346个氨基酸,牦牛KLF3氨基酸序列与普通牛(XP_010820342.1)的氨基酸序列同源性达100%。KLF3 m RNA在牦牛肺脏和肝脏的表达水平较高,极显著高于其他组织(P<0.01)。

The aims of this study are to clone yak KLF3 gene sequence,to analyze its biological characterization,and to clarifyits expression patterns in different tissues. Six healthy,4-6 years old male Maiwa yaks were selected as experiment animals. After fastingslaughter,the tissue samples of spleen,lung,kidney,subcutaneous fat and longissimus dorsi muscle were collected for the total RNAextraction. The Reverse Transcription PCR（RT-PCR）was used to clone Maiwa yak KLF3 gene sequence,the quantitative real-time PCR（q PCR）was used to detect the expression levels of KLF3 gene in different tissues. The results showed that the sequence of yak KLF3 gene was 1 137 bp（Gen Bank accession number ：KX964630）,containing 963 bp CDS,32 bp 5＇ UTR and 64 bp 3＇ UTR,encoding 346 amino acids. The aminoacid sequence of yak KLF3 shared a similarity of 100% with that of cattle（XP_010820342.1）. The mRNA expression level of KLF3 was highin lung and liver tissue,significantly higher than other tissues（P 0.01）.