為了分析弓形蟲GT1蟲株GRA15（GRA15GT1）蛋白的反應原性，通過PCR擴增編碼GRA15GT1 52~635氨基酸肽段的基因片段，構(gòu)建pGEX-6P-1-GRA15GT1載體，轉(zhuǎn)化BL21菌誘導表達；通過SDS-PAGE和Western blot方法進行表達驗證及反應原性分析。結(jié)果顯示：SDS-PAGE及以GST標簽抗體為一抗進行Western blot，均有目的條帶，比理論值稍大；以豬弓形蟲陽性血清為一抗的Western blot條帶與GST抗體孵育后的條帶大小一致。上述結(jié)果表明，GRA15GT1蛋白具有較好的反應原性，這為下一步分段表達GRA15GT1蛋白，研究其在弓形蟲血清學分型中的應用奠定了基礎(chǔ)。
Expression and Immunoreactivity Exploration of GRA15Protein
of Toxoplasma gondii GT1 Strain
In order to analyze the immunoreactivity of GRA15protein of Toxoplasma gondii GT1 strain（GRA15GT1），the gene encoding a 584-residue peptide（from aa52 to aa635）was amplified by PCR，then the pGEX-6P-1-GRA15GT1 vector wasconstructed and transformed into BL21 strain of E. coli. After inducibleexpression，the purified GRA15 protein were obtainedand verified by SDS-PAGE and Western blot. Results showed that，the target strip was slightly larger thanits estimated size when the GST-tag mouse monoclonal antibody was used asprimary antibodies，as well as the swine anti-T. gondii positive serum. The resultsindicated GRA15GT1 protein had good immunoreactivity，which would lay a foundation for the next stepof GRA15GT1 protein expression and its application in serological typing ofToxoplasma gondii.
全文下載鏈接：
http://kns.cnki.net/kcms/detail/detail.aspx?dbcode=CJFD&filename=ZGDW201901020&dbname=CJFDTEMP
當前位置：