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莱茵衣藻BBS1基因RNAi载体的构建及应用

DOI: 10.13560/j.cnki.biotech.bull.1985.2015.07.016, PP. 109-114

Keywords: 莱茵衣藻,RNAi载体,BBS1基因,趋光性

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Abstract:

巴德-毕氏综合症(Bardet-Biedlsyndrome,BBS)是一种由多种基因突变造成、与原生纤毛功能缺失相关的疾病,包含12种致病基因,分别为BBS1-12。旨在通过对衣藻BBS1基因进行RNAi干扰来研究其在衣藻中的功能。首先搭建衣藻快捷简单的RNAi骨架载体;再用RT-PCR克隆莱茵衣藻BBS1基因的一段cDNA序列于中间载体pEASY-T1上;测序后通过两次酶切连接到RNAi骨架载体上,经菌液PCR、酶切和测序验证,成功构建BBS1基因的RNAi干涉载体。经基因枪介导法转化莱茵衣藻CC503,转基因衣藻具有明显不同于对照的表现型,趋光性发生了改变。

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