猪ROCK1基因克隆、序列分析及时空表达研究

doi:10.16431/j.cnki.1671-7236.2018.03.002

《中国畜牧兽医》 ›› 2018, Vol. 45 ›› Issue (3): 571-580.doi: 10.16431/j.cnki.1671-7236.2018.03.002

猪ROCK1基因克隆、序列分析及时空表达研究

冯小婷^1,2, 朱记平¹, 张蕊蕊², 李毅¹

1. 武汉生物工程学院应用生物技术研究中心, 湖北省病毒载体(基因治疗)工程技术研究中心, 武汉 430415;
2. 华中农业大学, 农业部猪遗传育种重点开放实验室, 农业动物遗传育种与繁殖教育部重点实验室, 武汉 430070

收稿日期:2017-07-11 出版日期:2018-03-20 发布日期:2018-03-22
通讯作者: 李毅 E-mail:johnli2668@hotmail.com
作者简介:冯小婷(1984-),女,湖北鄂州人,博士,讲师,研究方向:动物遗传育种与分子病毒学,E-mail:xiaotingfeng1984@163.com;朱记平(1985-),女,湖北咸宁人,博士,讲师,研究方向:预防兽医学,E-mail:jp_zhu732@126.com
基金资助:
湖北省教育厅科学技术研究计划指导性项目（B2016301）

Cloning, Sequence Analysis and Expression Patterns of Porcine ROCK1 Gene

FENG Xiaoting^1,2, ZHU Jiping¹, ZHANG Ruirui², LI Yi¹

1. Hubei Engineering Research Center of Viral Vector, Center for Applied Biotechnology, Wuhan Institute of Bioengineering, Wuhan 430415, China;
2. Key Laboratory of Agriculture Animal Genetics, Breeding and Reproduction, Ministry of Education, Key Laboratory of Swine Genetics and Breeding, Ministry of Agriculture, Huazhong Agricultural University, Whuan 430070, China

Received:2017-07-11 Online:2018-03-20 Published:2018-03-22

摘要/Abstract

摘要：

为进一步获得猪肉质性状候选基因Rho相关激酶1（Rho-associated，coiled-coil containing protein kinase 1，ROCK1）的序列特征、生物学功能及时空表达规律等信息，本研究通过PCR、SMARTer RACE PCR的方法克隆了猪ROCK1基因，应用生物信息学方法分析该基因的蛋白质序列在不同物种中的进化关系，并采用RT-PCR和实时荧光定量PCR的方法检测其在不同组织和不同猪种背最长肌不同发育阶段的表达。结果显示，ROCK1基因有2个转录本，包含4 065 bp的开放阅读框（ORF），编码1 354个氨基酸；猪ROCK1基因ORF序列与人、小鼠和大鼠的ORF序列的同源性分别为95%、91%和91%，相应的氨基酸序列同源性分别为98%、96%和94%，该蛋白在不同物种间高度保守；ROCK1基因的组织分布较广泛，不同发育阶段在同一组织中的表达会发生变化；胚胎期ROCK1基因在梅山猪背最长肌中的表达极显著高于大白猪（P < 0.01），出生后ROCK1基因在2个猪种背最长肌中的表达无显著差异（P > 0.05）。本试验结果为研究ROCK1基因在猪骨骼肌发育中的生物学功能及其分子机制奠定基础。

关键词: 猪; ROCK1基因; 克隆; 表达

Abstract:

To further obtain the information about the sequence structure,biology function and spatio-temporal expression of Rho-associated,coiled-coil containing protein kinase 1(ROCK1) gene for meat traits,ROCK1 gene was cloned through PCR and SMARTer RACE PCR,the sequence was analyzed by bioinformatics method,and the mRNA expression was detected by RT-PCR and quantitative Real-time PCR.The results showed that ROCK1 gene had two transcripts,both containd 4 065 bp open reading frame (ORF),encoding 1 354 amino acids,and the high sequence homology of ROCK1 gene among human,mice and rats were 95%,91% and 91%,while the corresponding amino acid sequence homology were 98%,96% and 94%,respectively.ROCK1 protein was highly conserved in different species.ROCK1 gene expressed widely in different tissues,and the expression level changed in varied development stages.The ROCK1 gene expression of embryonic longisimus dorsi in Meishan pigs was extremely significantly higher than that of Yorkshire pigs (P < 0.01),while there were no significant differences in other developmental stages after birth (P > 0.05).The results would provide a basis for further study of the biological function and mechanism of ROCK1 gene on porcine skeletal development.

Key words: porcine; ROCK1 gene; cloning; expression

中图分类号:

冯小婷, 朱记平, 张蕊蕊, 李毅. 猪ROCK1基因克隆、序列分析及时空表达研究[J]. 《中国畜牧兽医》, 2018, 45(3): 571-580.

FENG Xiaoting, ZHU Jiping, ZHANG Ruirui, LI Yi. Cloning, Sequence Analysis and Expression Patterns of Porcine ROCK1 Gene[J]. , 2018, 45(3): 571-580.

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猪ROCK1基因克隆、序列分析及时空表达研究

Cloning, Sequence Analysis and Expression Patterns of Porcine ROCK1 Gene

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