Cloning and expression characteristics of the pig Stra8 gene.
Cloning and expression characteristics of the pig Stra8 gene.
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猪Stra8基因的克隆及表达特征。
DOI：
10.3390/ijms150712480
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发表时间：
2014-07-15
影响因子：
5.6
通讯作者：
Zhang Y
中科院分区：
文献类型：
--
作者：
Wang X;Chen T;Song C;Gao B;Zhang Y
Stra 8（Stimulated by Retinoic Acid 8）被认为是减数分裂的看门基因。利用反转录PCR和RACE技术，克隆了猪Stra 8基因的全序列。对该序列进行了生物信息学分析。采用半定量方法，检测Stra 8在成年梅山猪和母猪睾丸、附睾尾、附睾体、附睾头、精囊、前列腺、考珀腺、心、肝、脾、肺、肾、胃、下丘脑、垂体、大脑、小脑和海马等组织中的表达特征。采用实时荧光定量PCR技术分析了2、30、60、90和150日龄梅山公猪睾丸中的表达模式。构建了Stra 8基因的真核表达载体，并将其转染体外培养的NIH-3 T3细胞和第三代猪精原干细胞（SSCs）。在不同时间点评价睾丸重量和附睾尾精子计数。结果表明，猪Stra 8基因cDNA长度为1444 bp，编码366个氨基酸，具有典型的螺旋-环-螺旋（HLH）结构域。这是一种测试特异性表达。在猪睾丸中的表达从第2天开始，并且随着年龄和体重的增加而显著增加（p < 0.05）。将真核表达载体EGFP（enhanced绿色fluorescent protein）-N1-pStra 8转染NIH-3 T3细胞和猪精原干细胞后，可在NIH-3 T3细胞的胞浆中表达。而在精原干细胞中，Stra 8主要表达于细胞质，细胞核中表达较少。我们的数据表明Stra 8可能作为转录因子启动幼猪减数分裂。
Stra8 (Stimulated by Retinoic Acid 8) is considered a meiotic gatekeeper gene. Using reverse transcriptase PCR and rapid amplification of cDNA ends (RACE), the complete sequence of the pig Stra8 gene was cloned. Bioinformatics analyses of this sequence were performed. Using semi-quantitative methods, the expression characteristics of Stra8 in Testis, cauda epididymis, body epididymis, caput epididymis, seminal vesicles, prostate gland, Cowper’s gland, heart, liver, spleen, lung, kidney, stomach, hypothalamus, pituitary gland, cerebrum, cerebellum, and hippocampus of adult Meishan boar and sow tissues were examined. The expression pattern in the testis of 2-, 30-, 60-, 90-, and 150-day old Meishan boars were analyzed using real-time PCR. We constructed a eukaryotic expression vector for the Stra8 gene and used it to transfect NIH-3T3 cells and third generation pig spermatogonial stem cells (SSCs) cultured in vitro. Testes weight and sperm count in the cauda epididymis were evaluated at various time points. The results showed that the length of the pig Stra8 gene cDNA was 1444 bp encoding 366 amino acids with one typical helix-loop-helix (HLH) domain. It is testes-specific expression. Expression was first detected in boar testis starting at day 2, and its expression significantly (p < 0.05) increased with age and body weight. When NIH-3T3 cells and pig SSCs were transfected with the eukaryotic expression vector EGFP (enhanced green fluorescent protein)-N1-pStra8, it was expressed in the cytoplasm of NIH-3T3 cells. However, in SSCs, Stra8 was expressed predominantly in cytoplasm and few in nucleus. Our data suggest that perhaps Stra8 acts as a transcription factor to initiate meiosis in young boar.
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影响因子：
56.9
作者：
Bowles, J;Knight, D;Koopman, P
通讯作者：
Koopman, P
影响因子：
2.7
作者：
BOUILLET, P;OULADABDELGHANI, M;CHAMBON, P
通讯作者：
CHAMBON, P
影响因子：
3.6
作者：
Li, Hui;Palczewski, Krzysztof;Clagett-Dame, Margaret
通讯作者：
Clagett-Dame, Margaret
影响因子：
4.5
作者：
Bonache, S.;Algaba, F.;Larriba, S.
通讯作者：
Larriba, S.
DOI：
10.1073/pnas.0604871103
发表时间：
2006-10-17
影响因子：
11.1
作者：
Liu, Jie;Zheng, Qi;Lu, Min
通讯作者：
Lu, Min