梅山猪Tektin4基因的克隆及表达规律的研究

摘要/Abstract

摘要：

本研究旨在克隆猪Tektin4基因，并研究该基因在梅山猪组织器官中的时空表达规律。首先，采用RACE方法获得猪Tektin4的cDNA序列，对所获序列进行生物信息学分析；其次采用半定量的方法检测Tektin4在150日龄梅山公母猪组织器官中的表达特征；最后，采用荧光定量的方法分析该基因在2、30、60、90、150日龄段梅山公猪睾丸中的表达规律，并对不同日龄段梅山公猪附睾尾的精子计数。结果显示，猪Tektin4基因cDNA序列全长为1 500 bp，编码447个氨基酸。具有TEKTIN家族保守结构域，即4个Coiled-coils结构，在螺旋Helix2A和Helix2B之间存在TEKTIN家族特征的信号序列RPNVELCRD。猪Tektin4基因编码的氨基酸序列与牛和马的相似性最高为89%，进化距离最近，与热带爪蟾的相似性最低为62%，距离最远。Tektin4在成年公猪睾丸中表达丰度较高。在公猪的垂体和母猪的子宫角和输卵管中表达丰度较低，该基因于60 d的睾丸中开始表达，与附睾尾出现完整形态的精子时间相一致。150 d的表达量显著高于其他日龄(P<0.01)。该基因编码氨基酸作为精子尾部的结构蛋白与精子发生有关，并可能与部分组织或器官纤毛发生有关。

Abstract:

The objective of this study was to clone pig Tektin4 gene and analyze the tempospatial expression patterns in organs or tissues of Meishan pigs. Using RACE, Tektin4 cDNA was cloned and bioinformatics analysis was performed. The expression of Tektin4 mRNA in tissues and organs of 150-day-old Meishan boar and sow were detected by semi-quantitative methods.The expression pattern of Tektin4 in the testis of 2-, 30- , 60-, 90- and 150-day-old Meishan boars were analyzed by Real-time PCR. Sperm in cauda epididymal of Meishan boars at different development stages were counted. The results showed pig Tektin4 cDNA was 1 500 bp, encoding 447 amino acids, which had TEKTIN family conservative domain of four Coiled-coils. Between Helix2A and Helix2B spirals, the characterized sequences RPNVELCRD for TEKTIN family were present. Pig Tektin4 amino acids sequence showed 89% identity with cattle and horse and were in the same cluster by phylogenetic analysis, and it had the lowest similarity (62%) and the longest evolution distance between pig and tropical xenopus. Tektin4 expressed highly in testis and weakly in uterine horn and oviduct of sow and pituitary of boar. It began to express in testis on 60 days, which paralleled with emergency of sperm with complete shape in cauda epididymal. The expression level in 150 days testis was higher significantly (P<0.01) than those in other days testis. It implied that Tektin4 has the function of sperm tail structural protein, which may be involved in structure of cilia of some organs or tissues.

中图分类号:

S828
S813.3

王宵燕，宋成义，高波，李碧春，陈庭锋，何庆玲. 梅山猪Tektin4基因的克隆及表达规律的研究[J]. 畜牧兽医学报, doi: .

WANG Xiao-yan, SONG Cheng-yi, GAO Bo, LI Bi-chun, CHEN Ting-feng, HE Qing-ling. Cloning and Expression Patterns of Tektin4 Gene in Meishan Pig[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, doi: .

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参考文献

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