CLONING AND COMPARISON OF THE GENES ENCODING PREPROAPAMIN FROM THE VENOM OF 2 HONEYBEE AND 4 WASP SPECIES

2种蜜蜂和4种胡蜂蜂毒明肽基因比较(英文)

Preproapamin genes were amplified by RT-PCR from total RNA from the venom glands of 2 honeybee species, Apis mellifera, A. cerana cerana, and 4 wasp species, Vespa magnifica, V. velutina nigrothorax and Polistes hebraeus, respectively. Their PCR products were ligated into pGEM -T easy vector and the nucleotide sequences analyzed. The six fragments were all 141?bp in length and contained a n ORF coding the precursor of apamin. The apamin precursors of V. magnifica, V. velutina nigrothorax and P. hebraeus had 95% and 93% similarity with that of A. melliera in nucleotide and amino acid sequences, respectively. That of Vespu la maculifrons was identical to that of A. mellifera in nucleotide and amino acid sequences. Apamin precursors of V. magnifica, P. hebraeus and V. velutina nigrothorax also had the same nucleotide sequences. The nucleotide sequences o f preproapamin genes from the Chinese honeybee, A. cerana cerana and 4 wasp sp ecies are described for the first time. A notable discovery was that the wasps species had exactly same apamins as the honeybees despite the fact they belong to different insect families.

从中华蜜蜂、意大利蜜蜂和大胡蜂、墨胸胡蜂、额斑黄胡蜂、亚非马蜂 6种蜂的雌成蜂毒腺中快速抽提总RNA ,用RT PCR方法分别扩增各得到大小约为 15 0bp的cDNA片段 ,进一步将这 6个片段克隆入pGEM Teasy载体 ,进行测序和序列分析。结果表明 ,所扩增得到的 6个片段长度均为 14 1bp ,均为编码蜂毒apamin前体的cDNA。序列比较发现 ,中华蜜蜂、大胡蜂、墨胸胡蜂和亚非马蜂序列完全一样 ,都与已发表意大利蜜蜂precursorofapamin核苷酸序列具有 95 %同源性 ,氨基酸序列具有 95 %的同源性。额斑黄胡蜂和意大利蜜蜂precursorofapamin核苷酸序列完全一样。本研究首次从中华蜜蜂与几种胡蜂中克隆到蜂毒precursorofapamin基因 ,并发现胡蜂科与蜜蜂科来源的蜂毒明肽序列是完全保守的。