松墨天牛幼虫cDNA文库的构建及EST分析

cDNA Library Construction and EST Analysis of Monlchamus alternatus Larvae

松墨天牛（Monlchamus alternatus Hope）是林业重大害虫。本研究运用SMART技术构建了松墨天牛幼虫cDNA文库,库容量为1.09×10^6,文库滴度为5.45×10^6 pfu/mL,重组率为97.8%,平均插入片段大小约为1 000 bp,构建的文库质量较好。随机选取1 022个克隆进行测序,获得了1 012条表达序列标签（ESTs）,序列在GenBank中登录号为JZ143720～JZ144731。EST序列经过聚类拼接得到411条非重复单一序列,其中216条序列有功能注释。在216条有功能注释的序列中,有181条（83.80%）参与＂分子功能＂,122条（56.48%）参与＂细胞成分＂,168条参与＂分子生物学过程＂（77.78%）。研究结果为克隆和分析松墨天牛新的功能基因奠定了基础。

Monochamus alternatus（M.alternatus） Hope is a serious pest management concern in stands of pines.In this study,a cDNA library of the M.alternatus larva was constructed by SMART（switching mechanish at 5＇ end of RNA transcript） technology.The results showed that the numbers of depent clones of first was 1.09 ×10^6,the titer was 5.45×10^6pfu/mL,the recombination rate was 97.8%,and the average size of inserted cDNA fragments was about 1 000 bp.A total of 1 012 expressed sequence tags（ESTs）（GenBank accession No.：JZ143720～JZ144731）were generated successfully after sequencing,and 411 unique expressed sequences were obtained after cluster analysis.Among the 411 unique expressed sequences,216 ESTs could be functionally annotated,of which 181 ESTs（83.80%） participate in the＂molecular function＂,122 ESTs（56.48%） participate in the＂cellular component＂,and168 ESTs（77.78%） participate in＂biological process＂.This study may benefit for the further research on the molecular biology of M.alternatus,and lay a foundation for cloning and analyzing the functions of M.alternatus genes.