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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,構建pGEX-6P-1-GRA15GT1載體,轉化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。
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.
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