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Int. J. Mol. Sci. 2014, 15(7), 12480-12494; doi:10.3390/ijms150712480

Cloning and Expression Characteristics of the Pig Stra8 Gene

†,* ,
Received: 19 May 2014 / Revised: 16 June 2014 / Accepted: 16 June 2014 / Published: 15 July 2014
(This article belongs to the Section Biochemistry, Molecular Biology and Biophysics)
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Abstract: 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.
Keywords: Stra8; Meishan pig; expression Stra8; Meishan pig; expression
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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MDPI and ACS Style

Wang, X.; Chen, T.; Song, C.; Gao, B.; Zhang, Y. Cloning and Expression Characteristics of the Pig Stra8 Gene. Int. J. Mol. Sci. 2014, 15, 12480-12494.

AMA Style

Wang X, Chen T, Song C, Gao B, Zhang Y. Cloning and Expression Characteristics of the Pig Stra8 Gene. International Journal of Molecular Sciences. 2014; 15(7):12480-12494.

Chicago/Turabian Style

Wang, Xiaoyan; Chen, Tingfeng; Song, Chengyi; Gao, Bo; Zhang, Yani. 2014. "Cloning and Expression Characteristics of the Pig Stra8 Gene." Int. J. Mol. Sci. 15, no. 7: 12480-12494.

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