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

doi:10.16431/j.cnki.1671-7236.2015.07.003

›› 2015, Vol. 42 ›› Issue (7): 1640-1646.doi: 10.16431/j.cnki.1671-7236.2015.07.003

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

徐梦思, 黄涛, 刘丽娟, 杨飞, 鲁慧文, 翟腾蛟, 陈亚楠

石河子大学动物科技学院, 石河子 832000

收稿日期:2014-12-09 出版日期:2015-07-20 发布日期:2015-07-23
通讯作者: 黄涛 E-mail:taohuang100@sina.com
作者简介:徐梦思(1990-),女,新疆石河子人,硕士,研究方向:猪分子遗传育种,E-mail:1453855240@qq.com
基金资助:
国家自然科学基金项目(30901014、31060295)

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

XU Meng-si, HUANG Tao, LIU Li-juan, YANG Fei, LU Hui-wen, ZHAI Teng-jiao, CHEN Ya-nan

College of Animal Science and Technology, Shihezi University, Shihezi 832000, China

Received:2014-12-09 Online:2015-07-20 Published:2015-07-23

摘要/Abstract

摘要： 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492 bp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。

关键词: 猪; SKP1基因; 卵泡; 表达量

Abstract: 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 expression 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%,respectively.SKP1 mRNA was expressed in all tested tissues (muscle,fat,heart,liver,spleen,lung,kidney,stomach,duodenum,ovarian,tubal,uterine,uterine horn,pituitary,corpus luteum,brain,hypothalamus),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),respectively 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 development.

Key words: porcine; SKP1 gene; follicle; expression

中图分类号:

徐梦思, 黄涛, 刘丽娟, 杨飞, 鲁慧文, 翟腾蛟, 陈亚楠. 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[J]. , 2015, 42(7): 1640-1646.

XU Meng-si, HUANG Tao, LIU Li-juan, YANG Fei, LU Hui-wen, ZHAI Teng-jiao, CHEN Ya-nan. Cloning of Porcine SKP1 Gene and it's Expression Studies between Meishan and Duroc Pigs with Follicle[J]. , 2015, 42(7): 1640-1646.

参考文献

[1] Bai C,Sen P,Hofmann K,et al.SKP1 connects cell cycle regulators to the ubiquitin proteolysis machinery through a novel motif,the F-box[J].Cell,1996,86(2):263-274.
[2] Connely C,Hieter P,Budding Y.Budding yeast SKP1 encodes an evolutionarily conserved kinetochore protein required for cell cycle progression[J].Cell,1996,86(2):275-285.
[3] 赵志超.梅山与杜洛克母猪卵泡间差异表达基因的分离研究[D].石河子:石河子大学,2012.
[4] Lingelbach L B,Kaplan K B.The interaction between Sgt1p and Skp1P is regulated by HSP90 chaperones and is required for proper CBF3 assembly[J].Molecular and Cellular Biology,2004,24(20):8938-8950.
[5] Liu F,Ni W,Griffith M E,et al.The ASK1 and ASK2 genes are essential for arabidopsis early development[J].The Plant Cell,2004,16(1):5-20.
[6] Piva R,Liu J,Chiarle R,et al.In vivo interference with SKP1 function leads to genetic instability and neoplastic transformation[J].Molecular and Cellular Biology,2002,22(23):8375-8387.
[7] 王宇峰.Skp1蛋白在小鼠早期胚胎发育过程中的功能研究[D].南京:南京医科大学,2008.
[8] Nagyova E.Regulation of cumulus expansion and hyaluronan synthesis in porcine oocyte-cumulus complexes during in vitro maturation[J].Endocrine Regulations,2012,46(4):225-235.
[9] Galloway S M,McNatty K P,Cambridge L M,et al.Mutations in an ooeyte-derived growth factor gene(BMP15) cause increased ovulation rate and infertility in a dosage-sensitive manner[J].Nature Genetics,2000,25(3):279-283.
[10] Kitagawa K,Abdulle R,Bansal P K,et al.Requirement of SKP1-Bubl interaction for kinetochore-mediated activation of the spindle checkpoint[J].Molecular Cell,2003,11(5):1201-1213.
[11] Seol J H,Shevchenko A,Deshaies R J.SKP1 forms multiple protein complexes,including RAVE,a regulator of V-ATPase assembly[J].Nature Cell Biology,2001,3(4):384-391.
[12] Hochstrasser M.Ubiquitin,protesaomes,and the regulation of intracellular protein degradation[J].Current Opinion in Cell Biology,1995,7(2):215-223.
[13] Varshaysky A.The ubiquitin system[J].Trends in Biochemical Sciences,1997,22:383-390.
[14] SCF^FBXL19调节Rac3蛋白泛素化和降解的研究[D].长春:吉林大学,2014.
[15] Hershko A.Ubiquitin:Roles in protein modification and breakdown[J].Cell,1983,34(1):11-12.
[16] DeSalle L M,Pagano M.Regulation of the G1 to S transition by the ubiquitin pathway[J].FEBS Letters,2001,490(3):179-189.
[17] Zheng N,Schulman B A,Song L,et al.Structure of the Cull-Rbxl-Skp1-FboxSkp2 SCF ubiquitin ligase complex[J].Nature,2002,416(6882):703-709.
[18] Feldman R M,Correll C C,Kaolan K B.A complex of Cdc4p,Skp1p,and Cde53p/cullin catalyzes ubiquitination of the phosphorylated CDK inhibitor Sic1p[J].Cell,1997,91(2):221-230.
[19] Mandel S A,Fishman-Jacob T,Youdim M B.Modeling sporadic Parkinson's disease by silencing the ubiquitin E3 ligase component,SKP1A[J].Parkinsonism Relat Disord,2009,15(3):148-151.
[20] 朱世平,孙丽,殷雪梅,等.胎次对杜洛克、长白和大白母猪繁殖性能的影响[J].中国畜牧兽医,2014,41(8):197-200.
[21] 黄涛,李大全,孙敬礼,等.猪排卵数性状研究进展[J].猪业科学,2010,27(11):92-94.
[22] 赵志超.梅山与杜洛克母猪卵泡间差异表达基因的分离研究[D].石河子:石河子大学,2012.

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

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