Septin14, a gene specifically expressed in the testis and seminal vesicle of the Banna mini-pig inbred line (BMI).
Septin14, a gene specifically expressed in the testis and seminal vesicle of the Banna mini-pig inbred line (BMI).
复制标题
Septin14，在版纳小型猪自交系（BMI）睾丸和精囊中特异表达的基因
DOI：
10.1590/1984-3143-ar2020-0521
复制
发表时间：
2021-02-16
影响因子：
1.7
通讯作者：
Huo J
中科院分区：
文献类型：
--
作者：
Wang P;Zhang X;Huo H;Wang S;Song X;Huo J
摘要Septin14是一个重要的精子发生相关基因，与男性不育的发病机制有关，但目前对其研究尚不深入。本研究利用RACE方法克隆了版纳小型猪近交系（BMI）Septin14全长cDNA，并在猪肾上皮细胞（PK15）和大肠杆菌中表达。coliRosetta（DE3）细胞。Septin 14在不同发育阶段的体细胞组织和睾丸中表达。猪Septin14 CDS长1，299 bp，编码432个氨基酸的肽（或蛋白质）（MW = 50.4 kDa）。系统进化分析表明，猪Septin14在进化上高度保守。GFP标记的Septin14融合蛋白的亚细胞定位显示，Septin14分布在整个睾丸细胞。在34份猪组织中，Septin14 mRNA在睾丸和精囊中特异表达。在6个不同的出生后发育阶段，睾丸Septin14 mRNA水平几乎检测不到的第2天，而最高水平出现在75天。本文首次报道了Septin14在猪中的时空表达谱，表明Septin14可能参与生殖细胞的分裂、发育和凋亡。利用pET原核表达系统，表达并分离了重组的67.9kDa Septin14蛋白。
Abstract Septin14 is an important spermatogenesis related gene involved in the pathogenesis of male infertility that has not been well studied. Here, full-length Septin14 cDNA of the Banna mini-pig inbred line (BMI) was cloned using the RACE method and expressed in pig kidney epithelial cells (PK15) and E. coli Rosetta (DE3) cells. Septin14 expression was identified in somatic tissues and testis in different developmental stages. The pig Septin14 CDS is 1,299 bp long, and encodes a peptide (or protein) of 432 amino acids (MW=50.4 kDa). Phylogenetic analysis indicated that pig Septin14 was highly evolutionarily conserved. Subcellular localization of GFP-tagged Septin14 fusion protein revealed that Septin14 was distributed throughout the testicular cells. Among 34 pig tissues, Septin14 mRNA was found specifically in testis and seminal vesicle. In six different postnatal developmental stages, the testicular level of Septin14 mRNA was barely detectable on day 2, while the highest level occurred on day 75. The spatiotemporal expression profile of Septin14, reported herein for the first time in pig, indicated that Septin14 might be involved in the division, development and apoptosis of germ cells. Furthermore, using a pET prokaryotic expression system, we expressed and isolated recombinant 67.9 kDa Septin14 protein.