Tektin3基因在猪生殖系统发育中的表达规律研究

Cloning and Expression of Tektin3 Gene in Swine Reproductive System Development

TEKTIN是附着于精子尾部基因丝的重要组分,对于基因丝的稳定和结构的复杂性起重要作用。Tektin3是其家族成员之一。研究采用RACE方法,获得梅山猪Tektin3基因全序列,利用生物信息学方法分析其结构和功能,采用半定量的方法研究150日龄梅山公母猪23个不同组织Tektin3基因的表达特征,并用Realtime PCR方法分析2、30、60、90和150日龄梅山公猪睾丸的表达规律。结果表明:(1)Tektin3基因全序列1 761bp,包含5'UTR 95bp,完整的CDS序列1 473bp和3'UTR 193bp,编码490个氨基酸,相对分子质量为56 481.3,理论等电点为7.94。(2)Tektin3基因在睾丸中高丰度表达,在下丘脑和子宫角中表达丰度较低。(3)Tektin3基因从梅山猪60日龄开始表达,到90日龄时显著下降(P<0.05),150日龄时显著上升(P<0.05)。结果表明该基因编码氨基酸除与精子发生有关,可能还与部分组织或器官的纤毛发生及公猪的性发育有关。

TEKTINS were essential for construction of axoneme in sperm flagella.Tektin3 is one member of TEKTINs family.This study aimed to study the expression of Tektin3 gene in Meishan pigs.The cDNA of Tektin3 gene was cloned with RACE to get its sequence and its structures were analyzed by bioinformatics.The mRNA distribution of the porcine Tektin3 was analyzed in 23 tissues of Meishan boars and sows with Semi-quantitative PCR.The expression of Tektin3 was analyzed in testis of 2-day,30-day-old,60-day-old,90-day-old,150-day-old Meishan boars with Realtime PCR.The result showed as follows:(1)The full length sequence of Tektin3 was 1 761 bp in length,containing 95 bp 5'UTR and coding region(CDS,1 473 bp)and 193 bp 3'UTR and encoding 490 amino acids.The isoelectric point(PI)of the protein was 7.94,and the molecular weight was 56 481.33.(2)The expression level of porcine Tektin3 was significant in testis but not significant in hypothalamus and uterinehorn.(3)Tektin3 begun to express in 60-day-old testis,but a significant drop in 90 days(P<0.05),a significant increase in 150 days(P<0.05).It implied that besides the function on sperm tail structural protein,the Tektin3 may be involved in structure of cilia of some organs or tissues and sexual maturity of boars.