生物技术通报 ›› 2017, Vol. 33 ›› Issue (8): 132-138.doi: 10.13560/j.cnki.biotech.bull.1985.2017-0204

• 研究报告 • 上一篇    下一篇

牦牛KLF3基因克隆及组织表达分析

林森1,2, 林亚秋1, 朱江江2, 柏雪1, 江明锋1, 王永1,2   

  1. 1. 西南民族大学生命科学与技术学院,成都 610041;
    2. 青藏高原动物遗传资源保护与利用四川省重点实验室,成都 610041
  • 收稿日期:2017-03-17 出版日期:2017-08-01 发布日期:2017-08-01
  • 作者简介:林森,男,硕士研究生,研究方向:动物遗传育种与繁殖;E-mail:ls2349@163.com
  • 基金资助:
    科技部科技支撑子项目(2014BAD13B03),中央高校基本科研业务费专项基金项目(2017NZYQN14)

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

LIN Sen1,2, LIN Ya-qiu1, ZHU Jiang-jiang2, BAI Xue1, JIANG Ming-feng1, WANG Yong1,2   

  1. 1. School of Life Science and TechnologySouthwest Minzu UniversityChengdu 610041;
    2. Key Laboratory of Sichuan Province for Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and ExploitationChengdu 610041
  • Received:2017-03-17 Published:2017-08-01 Online:2017-08-01

摘要: 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(qPCR)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(GenBank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 mRNA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)。

关键词: 牦牛, KLF3基因, 克隆, 组织表达

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

Key words: yak, KLF3 Gene, clone, tissue expression