沙子岭猪ASB17基因克隆、生物信息学分析及组织差异表达研究  

Cloning,Bioinfornatics and Tissue Differential Expression Analysis of ASB17 Gene in Shaziling Pig

作　　者：董莲花 冉茂良[1,2] 翁波[1,2] 李智[1,2] 彭馥芝 陈斌[1,2] DONG Lian-hua RAN Mao-liang WENG Bo LI Zhi PENG Fu-zhi CHEN Bin(College of Animal Science ＆ Technology , Hunan Agricultural University, Changsha 410128, China Hunan Provincial Key I~oratory for Genetic Improvement of Domestic Animal, Changsha 410128, China)

机构地区：[1]湖南农业大学动物科学技术学院,长沙410128 [2]畜禽遗传改良湖南省重点实验室,长沙410128

出　　处：《中国畜牧兽医》2016年第11期2811-2819,共9页China Animal Husbandry & Veterinary Medicine

基　　金：国家现代农业产业技术体系建设专项资金(CARS-36)

摘　　要：本研究利用RACE法克隆沙子岭猪ASB17基因cDNA全长并进行生物信息学分析,应用实时荧光定量PCR对ASB17基因在沙子岭猪D30时期9个不同组织(睾丸、肌肉、小肠、肺脏、心脏、肾脏、肝脏、脾脏、脂肪)及睾丸组织8个不同发育阶段(E90、D1、D30、D60、D90、D120、D150、D180)进行差异表达分析。结果显示,ASB17基因克隆所得片段长1 135bp,其中ORF区为888bp,编码295个氨基酸。实时荧光定量PCR结果显示沙子岭猪D30时期ASB17mRNA在睾丸中表达量最高,其次为脂肪及脾脏,而在肌肉及小肠中表达量最低,其中睾丸与肌肉、小肠及肺脏中表达量差异显著(P<0.05);睾丸组织不同发育时期中,ASB17mRNA在D120、D150时期表达量达到最高,在D180时期表达量略有下降,其中E90、D1、D30、D60与D90、D120、D150、D180时期差异极显著(P<0.01);D90与D180时期差异不显著(P>0.05),与其余时期均差异极显著(P<0.01);沙子岭猪性成熟期D120、D150、D180 3个时期表达量差异不显著(P>0.05)。本试验成功克隆了沙子岭猪ASB17基因cDNA全长并检测了其在猪不同组织及睾丸组织不同发育阶段表达量,为进一步研究ASB17基因功能奠定基础。In this study,ASB17 gene cDNA full length in Shaziling pig was cloned by RACE and it was analyzed by bioinformatics,the ASB17 gene expression levels were detected in Shaziling pig D30 period of nine different tissues（testis,muscle,intestine,lung,heart,kidney,liver,spleen,fat）and eight different developmental stages（E90,D1,D30,D60,D90,D120,D150,D180）of testicular tissue by Real-time quantitative PCR.The results showed that the length of ASB17 gene cDNA was 1 135 bp,and the ORF region was 888 bp,which encoded 295 amino acids.Real-time quantitative PCR results showed that the D30period＇s expression level of ASB17 gene mRNA was the highest in testis,followed by fat and spleen,while the expression of ASB17 gene mRNA in muscle and small intestine tissue were the lowest,there were significant difference in testis and muscle,small intestine,lung（P〈0.05）;In the testis,the expression of ASB17 gene mRNA was the highest in D120 and D150periods,while slightly declined in D180,the differences were extremely significant beween E90,D1,D30,D60 and D90,D120,D150,D180（P〈0.01）;The differences were not significant beween D90 and D180（P〈0.05）,while there were extremely significant with oth-er periods（P〈0.01）;The sexual maturity periods of D120,D150,D180 had no significant differences（P〈0.05）.The full length of ASB17 cDNA was cloned and the expression levels of ASB17 gene in different tissues and different developmental stages were detected in this study,which laid the foundation for further research on the function of ASB17 gene.

关 键 词：沙子岭猪 ASB17基因 克隆 基因表达 

分 类 号：Q78[生物学—分子生物学]