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家蚕BmNaPi基因的克隆表达与酵母穿梭表达质粒的构建

DOI: 10.7668/hbnxb.2011.03.011, PP. 51-55

Keywords: 家蚕,BmNaPi,克隆,组织表达,酵母穿梭表达质粒

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Abstract:

磷酸盐是蚕必需的无机物,钠离子依赖性的磷酸盐转运蛋白(Na(+)-dependentphosphatecotransporter,NaPi)调节磷酸盐在生物体内的代谢和转运.本研究通过生物信息学分析和分子生物学试验获得了家蚕NaPi的同源基因BmNaPi,该基因位于家蚕8号染色体,编码区长1437bp,有10个外显子,编码478个氨基酸,预测蛋白序列在氨基末端含有一信号肽,序列中间有8个转膜结构域,与登录号为AAF7968、XP_001602874、EAA00281、XP-39379、XP00190194等同源蛋白的一致性均在70%及以上.RT-PCR检测该基因在龄3d家蚕幼虫9种组织中的血液、体壁、生殖腺和头中表达,而中肠、丝腺、马氏管和脂肪体中没有表达.最后成功构建了该基因的酵母穿梭表达质粒pGBKT7-BmNaPi.

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