家蚕Bmo-miR-2739对丝素重链基因Fib-H表达的调控作用  

Expressional Regulation of Bombyx mori Fibroin Heavy-chain Gene by Bmo-miR-2739

作　　者：宋菲[1] 王欣[1] 钱平[1,2] 唐顺明[1,2] 赵巧玲[1,2] 郭锡杰[1,2] 沈兴家[1,2] 

机构地区：[1]江苏科技大学,江苏镇江212018 [2]中国农业科学院蚕业研究所,农业部蚕桑遗传改良重点实验室,江苏镇江212018

出　　处：《蚕业科学》2014年第3期404-408,共5页ACTA SERICOLOGICA SINICA

基　　金：国家重点基础研究发展计划"973"项目(No.2012CB1146-05);国家自然科学基金项目(No.31172266)

摘　　要：MicroRNAs(miRNAs)是一类内源性的非编码小RNAs,可通过序列互补配对的方式对靶基因mRNA的转录进行调控。丝素重链(Fib-H)是家蚕丝蛋白的重要组成成分,研究miRNAs对Fib-H基因mRNA的调控作用,有助于理解丝腺高效合成蚕丝蛋白的分子机制。用靶基因预测软件RNA22和RNAhybrid对Fib-H基因的3'-UTR及已知的家蚕miRNA进行分析,发现Fib-H基因的3'-UTR有一个Bmo-miR-2739的潜在靶位点。进一步将人工合成的Bmo-miR-2739模拟物和带有Fib-H基因3'-UTR的荧光素酶报告基因载体共转染BmN细胞,48 h后检测到荧光素酶表达上调约13%。初步推测Bmo-miR-2739对Fib-H具有正调控作用。MicroRNAs （miRNAs） are a class of endogenous non-protein coding small RNAs that regulate expression of target genes at post-transcriptional levels by base pairing with target mRNAs. Silk fibroin heavy chain （Fib-H） is an im- portant component of the silk protein in silkworm （ Bombyx mori）, To study the regulatory function of miRNAs on Fib-H gene will provide a better understanding of the molecular mechanism of high efficiency synthesis of silk protein in silk gland. We analyzed the 3＇-UTR of Fib-H gene and the known miRNAs of silkworm by using target gene prediction soft- ware RNA22 and RNAhybrid, and found a target site of Bmo-miR-2739 in the 3＇-UTR of Fib-H gene. To verify the regula- tory function of Bmo-miR-2739 on the expression of Fib-H gene, Fib-H 3＇-UTR was fused with firefly luciferase gene to construct the reporter plasmid, Then, BmN cells were co-transfected with artificially synthesized Bmo-miR-2739 mimics and the reporter plasmid. The result indicated that luciferase expression was up-regulated by 13% at 48 h after transfec- tion, suggesting that Bmo-miR-2739 has positive regulatory function on Fib-H.

关 键 词：家蚕 MICRORNA 丝素重链基因 转录调控 

分 类 号：S881.2[农业科学—特种经济动物饲养] Q522.5[农业科学—畜牧兽医]