猪SKP1基因的克隆及其在梅山猪与杜洛克猪卵泡中的表达研究

Cloning of Porcine SKP1 Gene and it's Expression Studies between Meishan and Duroc Pigs with Follicle

为探索SKP1(S-phase kinase association protein 1)基因在猪卵泡中的表达规律,本试验从猪卵泡组织中克隆了猪SKP1基因CDS区全长序列,采用Real-time PCR方法检测SKP1基因在不同组织中的表达谱,进一步分析了该基因在梅山猪和杜洛克猪S卵泡、M1卵泡、M2卵泡、L卵泡中的表达。结果表明,经克隆测序,得到了猪SKP1基因492bp编码区全长序列,与羊、人、黑猩猩、牛、大鼠的同源性分别为93.10%、92.90%、92.29%、91.89%、89.86%。SKP1基因在各组织中均有不同程度的表达(肌肉、脂肪、心脏、肝脏、脾脏、肺脏、肾脏、胃、十二指肠、卵巢、输卵管、子宫、子宫角、垂体、黄体、大脑、下丘脑),其中在子宫、脾脏、输卵管中表达量较高。SKP1基因的表达量在梅山猪S卵泡、M1卵泡和M2卵泡中的表达量均高于杜洛克猪,特别是在梅山猪M1卵泡和M2卵泡中SKP1基因的表达量极显著高于杜洛克猪(P<0.01),分别达到了2.39、2.82倍,而在L卵泡中的表达量却是杜洛克猪高于梅山猪,结果提示SKP1基因可能参与猪卵泡发育过程。

To detect the expression pattern of S-phase kinase association protein 1 （SKP1） gene in during porcine follicle development. The coding sequence （CDS） of SKP1 gene was cloned from porcine follicular tissue. To analyze the tissue expression profiling of SKP1 gene and its expres- sion level in follicles with different diameter （S, M1, M2, L） from Duroc and Meishan pigs were investigated by Real-time fluorescence quantitative PCR. The results showed that the length of CDS of porcine SKP1 was 492 bp,and it＇s similarity with that of Ovis aries, Homo sapiens, Pan troglodytes ,Bos taurus and Rattus were 93.10%,92.90% ,92.29% ,91.89% and 89.86% ,respec- tively. SKP1 mRNA was expressed in all tested tissues （muscle,fat,heart,liver,spleen,lung,kid- ney, stomach, duodenum, ovarian, tubal, uterine, uterine horn, pituitary, corpus luteum, brain, hy- pothalamus） ,and especially high in uterine,spleen and tubal. Both in Meishan pig. The expression of SKP1 mRNA in S, M1 and M2 follicles were higher than Duroc pig. Especially, the expression in Meishan pig M1 and M2 follicles were significantly higher than Duroc pig （P〈0.01） ,respec- tively 2.39,2.82 times, and the expression of L follicle in Duroc pig was higher than Meishan pig, which suggested that SKP1 gene might be involved in the process of procine＇s follicular develop- ment.