title: Cloning and expression characteristics of the pig Stra8 gene.
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.
authors.affiliation: College of Animal Science & Technology, Yangzhou University, Yangzhou 225009, China. wxyan@yzu.edu.cn.
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: Wang
authors.affiliation: College of Animal Science & Technology, Yangzhou University, Yangzhou 225009, China. ctf20070702664@126.com.
authors.forename: Tingfeng
authors.initials: T
authors.lastname: Chen
authors.affiliation: College of Animal Science & Technology, Yangzhou University, Yangzhou 225009, China. cysong@yzu.edu.cn.
authors.forename: Chengyi
authors.initials: C
authors.lastname: Song
authors.affiliation: College of Animal Science & Technology, Yangzhou University, Yangzhou 225009, China. bgao@yzu.edu.cn.
authors.forename: Bo
authors.initials: B
authors.lastname: Gao
authors.affiliation: College of Animal Science & Technology, Yangzhou University, Yangzhou 225009, China. ynzhang@yzu.edu.cn.
authors.forename: Yani
authors.initials: Y
authors.lastname: Zhang
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D018507:Gene Expression Regulation, Developmental; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D041681:NIH 3T3 Cells; D009928:Organ Specificity; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D012737:Sex Factors; D013552:Swine; D013737:Testis / Q000254:growth & development / Q000378:metabolism
chemical_list: D048868:Adaptor Proteins, Signal Transducing