家蚕BmST2基因的克隆及转录活性检测.pdfVIP

生物技术通报 # # BIO TE CHN OLOG Y B ULLETIN 20 11 1 BmST2 孔卫青 杨金宏 ( , 725000) : 通过生物信息学分析和生物学试验获得了家蚕糖转运蛋白基因 Bm ST2 ( G enBank 登 号: GQ 87 1755), 基 因位于家蚕 27 号染色体, 开放阅读框 ( ORF) 长 1 398 bp, 编码 465 个氨基酸, 预测蛋白序列有典型的 Sug a _t 结构域和 11 个疏水的跨膜结构域, 与家蚕 Bm ST 1蛋白相似性和一致性分别达 79% 和 64% , 与登 号为 EAT 47626ED S35465 EAA11457和 EFA05337 的同源蛋白相似性在 50% 以上RT-PCR 检测基因在 5 龄第 3 天家蚕 幼虫的 9种组织中转 活 性, 结果显示, BmST 2基因除在脂肪体没有表达外, 其他组织均有表达最后成功构建了基因的酵母穿梭表达质粒 pG- BKT7-Bm ST2 : 家蚕 糖转运蛋白基因 Bm ST 2 克隆 序列分析 转 活性 Cloning and TranscriptionalActivity Determination ofBmST2 Gene from Silkworm(Bom byx mori) KongW e iqing Y ang Jinhong (TheK ey SericulturalLaboratory of Shaanxi, A nkang University, A nkang 725000) Abstract: Bm ST 3 gene( G enBank accession numbe : GQ 87 1755), located on s ilkwo m 27th ch om osome, w as obta ined th ough b io infom atics ana lys is and m o lecula b io log ica l expe mi ent. The OR F is 1 398 bp long, and encodes 465 am ino ac ids, w ith typical Sug- a _t dom a in and 11 hyd ophobic t ansm enb ane dom a ins ins ide. A com pa ison of BmST2 w ith BmST 1 f om s ilkwo m, evea led that the i am ino ac ids sha e 79% smi ila ity and 64% identity, and the compa ison of BmST2 w ith hom o logous p ote in EAT 47626, EDS35465, EAA 11457, EFA 05337 f om othe sou ces evealed that the i am ino ac ids sha e above 50% smi ila ity. It is shown by RT- PCR expe mi ents tha t BmST2 gene has exp ession in all the 9 tissues de tected on day 3 of the fifth insta la vae but fa t body. Then yeast shuttling exp ession plasm id pGBKT7-BmST 2 w as successfully const ucted. K ey w ords: Bombyx mori Suga t anspo te Bm ST2 gene Clone Seque