A SHUTTLE VECTOR WHICH FACILITATES THE EXPRESSION OF TRANSFECTED GENES IN TRYPANOSOMA-CRUZI AND LEISHMANIA
A SHUTTLE VECTOR WHICH FACILITATES THE EXPRESSION OF TRANSFECTED GENES IN TRYPANOSOMA-CRUZI AND LEISHMANIA
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DOI：
10.1093/nar/20.15.3963
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发表时间：
1992-08-11
影响因子：
14.9
通讯作者：
KENDALL, G
中科院分区：
文献类型：
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作者：
KELLY, JM;WARD, HM;KENDALL, G
克氏锥虫表达载体已经使用衍生自甘油醛3-磷酸脱氢酶（gGAPDH）基因的侧翼区的序列构建。新霉素磷酸转移酶（neo（r））基因作为选择性标记并入。使用电穿孔，我们已经将该载体引入克氏锥虫和利什曼原虫细胞中并赋予G418抗性。转化由载体的头-尾串联重复序列组成的大的染色体外环状元件介导。转化的表型在不存在G418的情况下稳定至少6个月，并且可以在通过克氏锥虫生命周期的传代期间维持。插入到载体（pTEX）内的表达位点中的外源基因可以在转化细胞中以高水平表达。据我们所知，本文描述了第一个锥虫穿梭载体和第一个载体，锥虫和利什曼原虫的功能。
A Trypanosoma cruzi expression vector has been constructed using sequences derived from the flanking regions of the glyceraldehyde 3-phosphate dehydrogenase (gGAPDH) genes. The neomycin phosphotransferase (neo(r)) gene was incorporated as a selectable marker. Using electroporation we have introduced this vector into both T.cruzi and Leishmania cells and conferred G418 resistance. Transformation is mediated by large extrachromosomal circular elements composed of head-to-tail tandem repeats of the vector. The transformed phenotype is stable for at least 6 months in the absence of G418 and can be maintained during passage through the T.cruzi life-cycle. Foreign genes inserted into an expression site within the vector (pTEX) can be expressed at high levels in transformed cells. To our knowledge this paper describes the first trypanosome shuttle vector and the first vector which functions in both trypanosomes and Leishmania.