多房棘球绦虫原头节cDNA表达文库的构建及初步鉴定
李淑萍;陈雅棠
【期刊名称】《中华医学杂志（英文版）》 【年(卷),期】2001(114)002
【摘要】Objective To construct a λgt11 cDNA expression library of Echinococcus multilocularis protoscolex isolated in China. rnMethods Echinococcus multilocularis protoscolex mRNA was extracted using a Quickprep MicromRNA purification kit based on combining of the disruptive and protective properties of guanidinium thiocyanate (GTC) with the speed and selectivity of oligo (dT)-cellulose chromatography in a spum-column with some modification. Purified mRNA (1.8 μg) was submitted to reverse transcription using random hexamers ［pd(N6)］. The double-strand blunt-ended cDNAs were ligated with an EcoRI/NotI adaptor to form a cohesive EcoRI end. Subsequently the synthesized cDNA was inserted into vector λgt11 EcoRI arms. After being packaged in vitro, λgt11 was put to an infectious bacteria Echinococcus coli (E.coli) strain Y1090; the recombinants were screened by color selection. PCR amplification was performed to evaluate the size of insertion DNA fragments.rnResults The recombinant ratio was nearly 100% and approximately 1×106 clones could be derived from this λgt11 cDNA library. PCR results indicated that the insertion DNAs were about 1.48 kb. rnConclusions A λgt11 cDNA expression library consisting of a
多房棘球绦虫原头节cDNA表达文库的构建及初步鉴定
