CAJ | 학술논문

目的：克隆版纳微型猪近交系（ BMI）不育和可育公猪TDRP1基因，分析其序列及mRNA表达水平上的差异，预测其蛋白质功能，并检测该基因在可育公猪中的组织表达分布情况。方法以猪NM＿001198925序列为模板，设计特异引物，采用RT－PCR方法结合测序获得TDRP1的cDNA序列并进行生物信息学分析；采用半定量PCR方法检测TDRP1在不育和可育公猪睾丸中的表达规律，分析该基因在可育公猪17种组织中的表达特征。结果获得了BMI TDRP1基因的编码区序列（ GenBank登录号：KJ186786），生物信息学分析表明其编码186个氨基酸，蛋白质相对分子质量（ Mw）为20．49×103，等电点（ pI）为5．86，无信号肽，有94．1％的概率位于细胞核，含有1个亮氨酸富集的核输出信号。不同物种的氨基酸序列比对表明猪TDRP1与人、恒河猴、小鼠和大鼠等哺乳动物的TDRP1相似性在73％～83．2％之间，其中与人、恒河猴的相似性较高。 mRNA表达分析表明，TDRP1在BMI不育和可育公猪睾丸间表达水平差异无显著，在精囊腺和前列腺中高表达，在睾丸和小脑中中度表达，在大脑和肾脏中低表达，在其余组织中不表达。结论成功克隆了BMI TDRP1基因的全长编码区序列并发现了BMI特有的2个SNP位点；TDRP1基因在BMI不育和可育公猪间序列完全一致，睾丸mRNA表达水平差异无显著性，多组织转录谱分析表明该基因存在明显的组织差异表达现象，在精囊腺和前列腺中有较高表达量，为深入研究TDRP1基因在精子发生方面的作用奠定了基础。
목적：극륭판납미형저근교계（ BMI）불육화가육공저TDRP1기인，분석기서렬급mRNA표체수평상적차이，예측기단백질공능，병검측해기인재가육공저중적조직표체분포정황。방법이저NM＿001198925서렬위모판，설계특이인물，채용RT－PCR방법결합측서획득TDRP1적cDNA서렬병진행생물신식학분석；채용반정량PCR방법검측TDRP1재불육화가육공저고환중적표체규률，분석해기인재가육공저17충조직중적표체특정。결과획득료BMI TDRP1기인적편마구서렬（ GenBank등록호：KJ186786），생물신식학분석표명기편마186개안기산，단백질상대분자질량（ Mw）위20．49×103，등전점（ pI）위5．86，무신호태，유94．1％적개솔위우세포핵，함유1개량안산부집적핵수출신호。불동물충적안기산서렬비대표명저TDRP1여인、항하후、소서화대서등포유동물적TDRP1상사성재73％～83．2％지간，기중여인、항하후적상사성교고。 mRNA표체분석표명，TDRP1재BMI불육화가육공저고환간표체수평차이무현저，재정낭선화전렬선중고표체，재고환화소뇌중중도표체，재대뇌화신장중저표체，재기여조직중불표체。결론성공극륭료BMI TDRP1기인적전장편마구서렬병발현료BMI특유적2개SNP위점；TDRP1기인재BMI불육화가육공저간서렬완전일치，고환mRNA표체수평차이무현저성，다조직전록보분석표명해기인존재명현적조직차이표체현상，재정낭선화전렬선중유교고표체량，위심입연구TDRP1기인재정자발생방면적작용전정료기출。
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 sequen-cing 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 identi-ty 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 ex-pressed 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.