弓形虫ZS2株抗原基因的扩增及克隆

扩增弓形虫ZS2株P30抗原基因，构建PcDNA3—P30真核表达重组质粒。方法本文采用PCR技术，自行设计一对寡核苷酸引物（P1，P2），从弓形虫ZS2基因组DNA中特异扩增出编码P30抗原的基因片段。扩增的目的片段经纯化后用EcoRI和Hind双酶切后，克隆到真核表达质粒pcDNA3中，转化入大肠杆菌TG1，用氨共青霉素和PCR初筛，将PCR扩增阳性的重组子用EcoRI和Kind双酶切鉴定。结果从弓形虫ZS2株DNA中扩增出1025bP的P30基因，构建重组质粒PcDNA3—P30，酶切产物的大小分别与预期相符。结论成功地对弓形虫ZS2株P30基因进行体外扩增及构建真核表达重组质粒PcDNA3—P30，为重组P30抗原及核酸疫苗研究做好准备。

AMPLIFICATION AND CLONING OF THE MAJOR SURFACE ANTIGEN(P30)GENE OF ZS2 STRAIN OF TOXOPLASMA GONDII

Aim To amplify P30 antigenic gene of ZS2 Strain of Toxoplasma gondii,and to construct therecombinant eukaryotic expression plasmid of pcDNA3-P30. Methods The pair of primer P30 was designed byautor,and the gene fragment encoding sequence of P30 was amplified by polymerase chain reaction(PCR),thege nomic DNA of ZS2 Strain of Toxoplasma gondii as the template. After having been digested by EcoR l/Hind ,the fragments of the PCR products were cloned into pcDNA3 vector,then transferred into Escherichia Colt(E.coli) TGI.Having been screened by ampicillion--resistance and PCR amplification,the PCR positive recombinantswere identified with EcoR I/Hind digestion. Results The gene encoding P30 was successfully amplified fromthe genomic DNA of ZS2 Strain of Toxoplasma gondii by PCR,and the recombinant plasmid pcDNA--P30 wasconstructed. Conclusions It showed that the amplification of P30 gene in vitro and the construction ofrecombi nant expressing plasmid of pcDNA -- P30 were successfully, which provided the condition for the expression ofP30 gene.