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深圳水库区双脐螺rDNA基因片段克隆与序列分析

, PP. 267-271

Keywords: 深圳水库,双脐螺,rDNA,基因克隆,序列分析

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Abstract:

目的克隆深圳水库区双脐螺rDNA基因片段并进行序列分析。方法采集深圳水库排洪渠水体的双脐螺,提取基因组DNA,PCR方法扩增rDNA基因片段,纯化后与pMD18-T质粒连接,转化大肠埃希氏菌JM109,筛选阳性克隆;重组质粒经双酶切鉴定后,进行序列测定,并以BLAST和MEGA4软件分析序列特点。结果深圳水库区双脐螺rDNA基因扩增片段大小约为959bp;重组质粒经双酶切鉴定与预期结果相符;克隆的rDNA序列含有959个碱基,BLAST比对结果显示,rDNA克隆序列与GenBank中3株库恩双脐螺(B.kuhniana)序列的同源性为99%,与2株藁杆双脐螺(B.straminea)的同源性为98%,与1株中介双脐螺(B.intermedia)及2株亚马逊双脐螺(B.amazonica)的同源性介于96%~97%。应用邻位连接法(Neighbor-jioning,NJ)和最小进化法(MinimumEvolution,ME)2种方法构建系统发生树,深圳水库区双脐螺与3株B.kuhniana的遗传距离小,同属一个分支。结论成功克隆了深圳水库区双脐螺rDNA基因片段,其基因遗传特征与B.kuhniana双脐螺接近。

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