【摘要】 从墨胸胡蜂毒腺中提取总RNA,经Oligo(dT)磁珠纯化mRNA后,用AMV反转录酶合成第一链cDNA.完成第二链cDNA合成后,过柱纯化50bp以上的片段,与载体pUC118连接,转化E.coliJM109,建立了cDNA文库.得到的780个阳性克隆中,重组率达到98.5%.大部分插入片段在500～900bp.随机挑选10个短序列测序,一个为蜂毒激肽基因序列.更多还原

【Abstract】 Total RNA was extracted from venom sac of Vaspa velutina nigrithorax Buysson and purified by oligo dT magnet separation. The first strand cDNA was synthesized using AMV reverse transcriptase system, and then the second strand by DNA Polymerase I. The fragments longer than 50 bp were purified and ligated to pUC118. The cDNA library containing 780 clones was constructed, in which more than 98.5% clones are positive. Most of insert DNA were about 500-900 bp. Ten random clone sequences were gotten, one is vespakinin gene. The others contain entire ORF. The data showed that cDNA library had reasonably good quality.更多还原