中文摘要：

目的构建带有线虫组织特异启动子的含不同ployQ的THAP11转基因线虫载体，并制备转基因线虫，为进一步研究THAP11基因的功能提供基础。方法设计引物，以THAP11-pcDNA3．1his／mycB真核表达载体为模板，PCR法扩增含不同ployQ的THAP11基因，将其克隆至带有线虫组织特异性启动子的pFx-unc119（Gene MarkerdsRed）载体。酶切及测序鉴定正确后，用显微注射的方法制备THAPll转基因线虫，用荧光显微镜观察载体荧光表达并提取转基因线虫基因组DNA、RNA，PCR、RT—PCR方法检测其整合及表达情况。结果PCR扩增出含不同ployQ的THAP11-HIS片段，长度约1000bp，测序正确并构建pFx—uncl19（THAP11）载体，THAP11转基因线虫的载体在线虫体内稳定表达红色荧光，转基因线虫基因组内有相应的THAP11插入及转录。结论成功构建的THAP11转基因线虫的载体能稳定转染线虫，将有助于THAP11功能的进一步研究。

英文摘要：

Objective To construct a recombinant eukaryotic vectors expressing THAP11, which carried different polyQ,with specific promotor of Caenorhabditis elegans tissues, and to create transgenic Caenorhabditis elegans for the research of the function of THAP11 via microinjection. Methods THAP11 carrying different polyQ were cloned by PCR using THAP11-pcDNA3.1 his/mycB as template with a pair of primers with HIS tag. Then, the THAP11- HIS was cloned into the vector with specific promotor of Caenorhabditis elegans tissues unc119. Transgenic Caenorhabditis elegans were created by microinjection to observe the expressing of fuorescence,and extract the genome DNA and total RNA. The integration and the transcription of THAP11 were analyzed by PCR and RT-PCR. Results The PCR product of THAP11 -HIS carrying different polyQ were all about 1 000 bp in length, and the transgenic Caenorhabditis elegans vectors carrying THAP11-HIS with specific promoter unc-119 were constructed,which were sequenced correctly. The expressing of red fuorescence was stable in transgenic Caenorhabditis elegans, THAP11 was integrated in the genome of the transgenic Caenorhabditis elegans, and could be transcripted. Conclusion The THAP11 transgenic Caenorhabditis elegans can be constructed,which may be useful for the further research of the function of THAP11.