中华蜜蜂mrjp1 cDNA的克隆及其序列分析

构建中华蜜蜂(Apis cerana cerana)8日龄工蜂头部cDNA文库，利用中蜂基因组的mrjp3部分基因片段作为杂交探针，采用DIG标记筛选cDNA文库，获得mrjps阳性克隆120个；对阳性克隆进行PCR扩增和测序，通过NCBI的BLAST序列比对，获得12个与印度蜂(Apis cerana india)、西方蜜蜂(Apis mellifera L.)mrjp1基因同源的中蜂mtjp1 cDNA片段，并进一步对中华蜜蜂mrjp1的cDNA全序列进行测定和分析。序列比对分析表明，东方蜜蜂(Apis cerana)与西方蜜蜂mrjp1的cDNA序列相似性为93．78％，中华蜜蜂与印度蜂的相似性高达99．36％，这一结果从分子水平证实中华蜜蜂与印度蜂有较近的共同祖先，而东方蜜蜂与西方蜜蜂的亲缘关系较远。

Cloning and Sequence Analysis of mrjp1 cDNA from Apis cerana cerana

A cDNA library was constructed from eight-day-old worker heads of Apis cerana cerana.A probe derived from part of Apis cerana genomic mrjp3 segment was used to screen this library.A total of 120 positive clones of mrjps were screened out from the cDNA library with DIG-probe.The positive clones were amplified and sequenced with T3/T7 primer.12 sequences were similar to Apis cerana india and Apis mellifera mrjp1 gene by BLAST analysis.The cDNA sequence analysis indicated that the homology was 93.78% between Apis mellifera mrjp1 and Apis cerana mrjp1 while the homology was 99.36% between Apis cerana cerana and Apis cerana india.This result confirmed in molecular level that Apis cerana cerana and Apis cerana india had a common ancestor since the relationship between Apis cerana and Apis mellifera was more far.