Molecular cloning, sequence characterization and mRNA tissue expression analysis of TDRP1 gene from the Banna minipig inbred line (BMI)
Received:June 21, 2014  
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KeyWord:TDRP1;Spermatogenesis;Banna minipig inbred line (BMI);Bioinformatics;Tissue expression patterns
王配 云南农业大学 云南省版纳微型猪近交系重点实验室, 昆明 ;云南农业大学 动物科学技术学院, 昆明
霍金龙 云南农业大学 云南省版纳微型猪近交系重点实验室, 昆明 ;云南农业大学 动物科学技术学院, 昆明
王淑燕 云南农业大学 云南省版纳微型猪近交系重点实验室, 昆明 ;云南农业大学 动物科学技术学院, 昆明
潘伟荣 云南农业大学 云南省版纳微型猪近交系重点实验室, 昆明 ;云南农业大学 动物科学技术学院, 昆明
查星琴 云南农业大学 云南省版纳微型猪近交系重点实验室, 昆明 ;云南农业大学 动物科学技术学院, 昆明
施晨 云南农业大学 云南省版纳微型猪近交系重点实验室, 昆明
曾养志 云南农业大学 云南省版纳微型猪近交系重点实验室, 昆明 ;云南农业大学 动物科学技术学院, 昆明
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      Objective To get TDRP1 gene of sterile and fertile boar of the Banna minipig inbred line (BMI), predict its function by bioinformatics analysis, and detect its expression patterns in the fertile boar. Methods Based on the NM_001198925 sequence, we designed specific primers and amplified BMI TDRP1 using RT-PCR method for sequencing and bioinformatics analysis. Meanwhile, the expression of TDRP1 in 17 organ tissues (heart, liver, spleen, lung, kidney, thymus, lymph nodes, skin, duodenum, stomach, cerebrum, cerebellum,testis, epididymis, seminal vesicle, prostate, and bulbourethral gland) of fertile BMI boar and in the testis of sterile and fertile BMI boars was analyzed by semi-quantitative RT-PCR. Results The experiment obtained 680 bp cDNA sequence (GenBank accession number: KJ186786) of BMI TDRP1, which encodes a protein of 186 amino acids with a predicted molecular weight (Mw) of 20.49 kDa and isoelectric point (pI) 5.86, and no signal peptide. It was a nuclear protein with a probability of 94.1% and had a leucine-rich nuclear export signals. Homology analysis of protein sequences revealed that BMI TDRP1 showed high identity with that of humans, macaca mulatta, mouse and rat. The RT-PCR analysis showed that TDRP1 had a similar expression in the testes of sterile and fertile BMI boars. It was highly abundant in the seminal vesicle and prostate, moderately expressed in cerebellum and testis and weakly expressed in cerebrum and kidney, while undetected in other 11 organ tissues. Conclusions We have cloned TDRP1 complete coding sequence, and found 2 SNPs,showing no difference in sequences and the testis mRNA expression levels between the fertile and sterile BMI boars. The multi-tissue transcription profile shows different expression levels in different organ tissues, being high in the seminal vesicle and prostate. The results of this study provide a foundation for further insight into the role of this gene in spermatogenesis.
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