首页|期刊导航|中华医学杂志（英文版）|Stage-specific expression genes of the Spirometra erinaceieuropaei plerocercoid screened by mRNA differential display technique
中华医学杂志（英文版）2004，Vol.117Issue(3)：366-370,5.
Stage-specific expression genes of the Spirometra erinaceieuropaei plerocercoid screened by mRNA differential display technique
Stage-specific expression genes of the Spirometra erinaceieuropaei plerocercoid screened by mRNA differential display technique
摘要
Abstract
Background The stage-specific expression of genes is one of the most characteristics of parasites. It has been found that a lot of genes of Spriometra erinaceieuropaei are specifically expressed in pleroceroid in large amount, but not expressed when the plerocercoid development into adult worm. The study is to screen other stage-specific ecpression genes of plerocercoid of Spirtmetra erinceieuropaei.Methods RNA was separately extracted by acid guanidinium thiocyanate-phenol-chloroform from plerocercoids and adult worms of Spirometra erinaceieuropaei, DNA contaminated in the RNA was digested by RNase-free DNase. After the RNA was reverse transcripted to cDNA using T12MA, T12MC, T12MG and T12MT anchor-primers, PCR was done using the same T12MN and one random primer with α 35S-dATP in the system. The PCR products were fractionated on an 8% denatured polyacrylamide gel. Differential bands of the plerocercoid found in the gel were cut out, amplified by PCR and sequenced. Northern hybridization was used to identify the stage-specific expression genes.Results Eleven differential bands were selected from the gel and classified into 3 kinds of gene fragments by hybridization, after they were amplified by PCR. Fragments 1 (238 bp) and 2 (383 bp), were confirmed by Northern hybridization, as being expressed in the plerocercoid. However, fragment 3 (433 bp), was expressed in both the plerocercoid and the adult worm. Data from the 3 gene fragments underwent homological analysis in GenBank. The sequence which was homologous with fragments 1 and 2 was not found, but fragment 3 had high homology with many kinds of 28S rRNA. Conclusions The gene expressions of plerocercoids are different from adult worms because they live in different hosts. Two types of different gene fragments from the plerocercoid were found by mRNA differential display technique.关键词
mRNA differential display/Spirometra erinaceieuropaei/PCR/stage-specific expressionKey words
mRNA differential display/Spirometra erinaceieuropaei/PCR/stage-specific expression分类
医药卫生引用本文复制引用
刘殿武,刘建波,张丽梅,王晓波..Stage-specific expression genes of the Spirometra erinaceieuropaei plerocercoid screened by mRNA differential display technique[J].中华医学杂志（英文版）,2004,117(3):366-370,5.基金项目
This work was supported by the Scientific and Technical Department of Hebei Province (No. 01245512D). （No. 01245512D）
