秀丽隐杆线虫全身性与组织特异性表达重组载体的构建

Construction of recombinant vectors introducing ubiquitous expression or tissue-specific expression in Caenorhabditis elegans

利用PCR技术从秀丽隐杆线虫(Caenorhabditis elegans)基因组DNA中扩增获得Act-1基因的核心启动子序列、T07F10基因的核心启动子序列与3UTR的Poly(A)信号序列以及从表达载体pEGFP-N1上扩增获得SV40早期mRNA的Poly(A)信号序列与EGFP基因.以克隆载体pBluescript SK+为基本骨架,分别构建由Act-1基因的核心启动子序列、T07F10基因的核心启动子序列调控的、EGFP作为报告基因的秀丽隐杆线虫全身性表达重组载体和组织特异性表达重组载体,为研究不同类型载体在秀丽隐杆线虫体内的表达模式提供基础.

To investigate the differences of recombinant vectors introducing ubiquitous expression and tissue specific expression in Caenorhabditis elegans, the core promoter region of Act-1 gene and T07F10 gene, and the Poly（A） signal region of T07F10 gene were amplified from the genomic DNA of C. elegans using PCR, and SV40 early mRNA sequence and EGFP gene were amplified from eukaryotic vector pEGFP-N1, respectively. Taking the cloning vector pBluescript SK＋ as backbone, four varied recombinant vectors were constructed, two of which, that is recombinant vectors pB-Pact-EGFP- T07F10T and pB-Pact-EGFP-SV40T, were expressed ubiquitously in C. elegans, regulated by the core promoter region of Act-1 gene, and the others, named as recombinant vectors pB-PT07F10-EGFP-T07F10T and pB-PT07F10-EGFP-SV40T, were expressed distinctively in specific tissues of C. elegans, regulated by the core promoter region of T07F10 gene. Further expression exploitation with these four recombinant vectors probably provided substantial information on different expression patterns using transgene technique in C. elegans.