摘要
旨在克隆鉴定牦牛lncFAM200B,为探索其在牦牛肌肉和脂肪发育过程中的调控作用奠定基础。以臀肌cDNA为模板,设计特异性引物扩增lncFAM200B全长序列,用在线软件CPC、CPAT对该lncRNA的编码能力进行预测,进一步通过原核表达试验鉴定其编码能力;采用实时荧光定量PCR(RT-qPCR)对lncFAM200B进行组织表达分析,通过TargetScan 7.1和miRanda预测与lncFAM200B具有潜在相互作用miRNA的靶基因,利用DAVID在线软件对靶基因进行GO和KEGG分析。结果显示,牦牛lncFAM200B长度为531 bp;相较于普通牛多59 bp,编码潜能为-1.2874,且原核表达试验验证该lncRNA不具备蛋白编码能力,表明牦牛lncFAM200B确为lncRNA;组织表达谱研究表明,lncFAM200B在牦牛肺脏组织中表达量较高。与lncFAM200B具有潜在相互作用miRNA的靶基因,如Sirt1、SCD5、KLF9、PTEN和MYPN等,与肌肉和脂肪发育相关。为进一步探讨牦牛lncFAM200B在肌肉和脂肪发育中的生物学功能提供参考依据。
The aim of this study was to clone and to identify the yak lncFAM200B,which might lay the foundation for exploring its regulatory role in yak muscle and fat during development.The primers for lncFAM200B gene were designed and used for the full-length cloning with the gluteal cDNA as template.The protein coding potential of lncFAM200B was predicted using the online prediction software CPC and CPAT,and then identified by prokaryotic expression assays.The tissue expression profile was analyzed by Real-time quantitative PCR(RT-qPCR),the target genes of miRNAs,which were interactions with lncFAM200B,were predicted by TargetScan 7.1 and miRanda.GO enrichment and KEGG pathway analysis were conducted with DAVID online software.The results showed that the full-length of yak lncFAM200B was 531 bp,which was 59 bp longer than that in cattle.Bioinformatics predicts results showed that its coding potential was-1.2874,and prokaryotic expression assay further confirmed that lncFAM200B had no protein coding ability,suggesting that yak lncFAM200B was a real lncRNA.Tissue expression profiling showed that the expression of lncFAM200B had a higher level in yak lung tissue.The lncFAM200B interaction miRNA targets analysis revealed that many genes,such as Sirt1,SCD5,KLF9,PTEN and MYPN,were related to muscle and fat development.Taken together,the present study laid the foundation for further research on the function and regulatory mechanism of yak lncFAM200B in muscle and fat development.
作者
赵丽玲
王会
柴志欣
王吉坤
王嘉博
武志娟
信金伟
钟金城
姬秋梅
ZHAO Liling;WANG Hui;CHAI Zhixin;WANG Jikun;WANG Jiabo;WU Zhijuan;XIN Jinwei;ZHONG Jincheng;JI Qiumei(Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization,Sichuan Province and Ministry of Education, Southwest Minzu University, Chengdu 610041, China;State Key Laboratory of Hulless Barley and Yak Germplasm Resources and Genetic Improvement, Lhasa 850000,China)
出处
《华北农学报》
CSCD
北大核心
2020年第5期220-230,共11页
Acta Agriculturae Boreali-Sinica
基金
西南民族大学研究生创新型科研项目(CX2019SZ77)
国家肉牛牦牛产业技术体系项目(CARS-37)
青藏高原生态畜牧业协同创新中心开放基金(QZGYXT02)
西藏自治区财政专项(XZNKY-2019-C-052)。
