家蚕黄酮合酶i基因bmfnsi的克隆与干涉载体的构建.pdfVIP

华北农学报# 2009, 24( 5) : 26- 30 I BmFNS I 孔卫青, 杨金宏, 彭云武, 胡必利 ( , 725000) : , , I NCBI I , EST BmFNS I 1 451 bp,7 345 , 179~ 295 2- e(II) RT-PCR 2 T7 dsRNA L4440 :; BmFNS I ; ; :S881.2 :A : 1000- 7091(2009) 05- 0026- 05 Cloning and Construction of RNAi Vector of Flavone Synthase I Gene, BmFNS I from Silkworm, Bombyx mori KONG We-i qing, YANG Jin-hong, PENG Yun-wu,HU Bi-li (The Key Sericultural Laboratory of Shaanxi,Ankang University,Ankang 725000, China) Abstract lavone iswidely existing in animals and plants.They have various physiological activity and the synthesiza- tion of them can be catalyzed by flavone synthase I gene.Using the reported amino acid sequences of other organisms and the scaffold and EST sequences of silkworm,silkworm flavone synthase I gene,BmFNS I ,was obtained by electronic clone. The gene is 1451bp in length and 7 exons in it by cloning and sequencing.The deduced protein contains 345 amino acids and a structural domain of 2-oxoglutarate (2OG) and e( II)-dependent oxygenase superfamily located in amino acids 179 and 295. It is shown by RT-PCR experiment that the gene expressed mostly in midgut in all the tissues tested.Then gene was subcloned to L4440,which has two T7 polymerase promoter in opposite orientation to synthesize dsRNA in vitro. Key words Bombyx mori; BmFNS I ; Clone;Tissue expression , 5, 4-[4- 10] I , , EST[ 11, 12] [ 1] [ 13] I( lavone synthase I, NS I) 2- , I (2OG-) e( II) , BmFNS I , [ 2] , BmFNS I , , L4440 :