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大白猪睾丸组织let-7c和miR-23a表达模式研究及表达载体构建
发表时间：2014-06-15 浏览量：2196 下载量：603
| 全部作者： | 周佳伟，叶莲芝，罗立凡，官凯锋，马昌萍，夏宣炎，雷彬，李凤娥 |
| 作者单位： | 华中农业大学农业部猪遗传育种重点开放实验室；华中农业大学理学院 |
| 摘 要： | 研究扩增克隆let-7c与miR-23a，并获得其前体及成熟微小RNA(micro RNA, miRNA)序列信息。利用RT-PCR方法检测let-7c，miR-23a在大白猪35，60，90，180日龄不同发育时期睾丸组织的表达情况。结果发现：pri-let-7c在180日龄表达水平最高，而在60日龄几乎不表达，但是成熟let-7c在180日龄睾丸中显著低于其他日龄（P<0.05）；pri-miR-23a在35日龄和90日龄表达量较高，而成熟miR-23a在90日龄表达量增加显著高于35日龄和180日龄（P<0.05）。通过RNAhybrid和RNA22预测到let-7c可能的靶基因精子粘附蛋白分子1（sperm adhesion molecule 1，SPAM-1）和miR-23a可能的靶基因TNF受体超家族6基因（TNF receptor superfamily member 6, FAS）；构建插入pri-miRNA片段的plet-7c，pmiR-23a表达载体进行猪睾丸（swine testicular，ST）细胞转染实验。应用RT-PCR方法进行表达验证，获得相应的表达量信息。 |
| 关 键 词： | 畜牧学；miRNAs；RT-PCR；猪；精子发生 |
| Title： | Expression patterns of let-7c and miR-23a in large white pig testes and miRNA expression vector construction |
| Author： | ZHOU Jiawei, YE Lianzhi, LUO Lifan, GUAN Kaifeng, MA Changping, XIA Xuanyan, LEI Bin, LI Feng’e |
| Organization： | Key Laboratory of Pig Genetics and Breeding of Ministry of Agriculture, Huazhong Agricultural University; College of Science, Huazhong Agricultural University |
| Abstract： | This study amplified and cloned let-7c and miR-23a, and obtained their precursor and mature sequences of miRNA. RT-PCR was used to analyze the expression pattern of let-7c and miR-23a in the porcine testes at age of 35, 60, 90, 180 d. The results showed that pri-let-7c had the highest expression at 180 d, but scarcely expressed at 60 d, while mature let-7c had a lower expression at 180 d than other age groups (P<0.05). pri-miR-23a highly expressed at 35 d and 90 d, while the mature miR-23a had a high expression at 90 d than at 35 d and 180 d (P<0.05). Using online RNAhybrid and RNA22, sperm adhesion molecule 1 (SPAM-1) and TNF receptor superfamily member 6 （FAS） genes were predicted to be the target genes of let7c and miR-23a, respectively. plet-7c and pmiR-23a expression vectors were constructed by inserting pri-miRNA fragment in, and then transfected pig swine testicular (ST) cells. RT-PCR was used for expression confirmation and obtained the corresponding expression information. |
| Key words： | animal husbandry; miRNAs; RT-PCR; pig; spermatogenesis |
| 发表期数： | 2014年6月第11期 |
| 引用格式： | 周佳伟，叶莲芝，罗立凡，等. 大白猪睾丸组织let-7c和miR-23a表达模式研究及表达载体构建[J]. 中国科技论文在线精品论文，2014，7（11）：1102-1107. |
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