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METHOD FOR DETERMINING THE SENSITIVITY OR RESISTANCE OF RETROVIRUS ISOLATES TO MOLECULES AND DIAGNOSTIC KITS

机译:测定反转录病毒分离株对分子和诊断试剂盒的敏感性或抗性的方法

摘要

A method of determining sensitivity or resistance of isolates of HIV retroviruses to a molecule includes a) amplifying sequences coding for a protease of a retrovirus to be studied, with or without the or some of amino acid sequences situated upstream and downstream of a cleavage site of a precursor in which the amino acid sequences are situated, b) recombining fragments of DNA, a final product of the amplification, and an expression vector allowing expression of sequence coding for the protease of the retrovirus to be studied under control of a known inducible promoter through co- transformation of the vector and the DNA fragments with at least one yeast cell, c) culturing co-transformed yeast cell or cells to obtain a sufficient number of transformants to perform a sensitivity or resistance test, and recovering transformants issuing from the co-transformed cell, on any suitable medium, d) incubating the transformants in the presence of a molecule to be tested, e) qualitatively or quantitatively analyzing the living cells, and f) deducing the sensitivity or resistance phenotype.
机译:确定HIV逆转录病毒分离株对分子的敏感性或抗性的方法包括:a)扩增编码要研究的逆转录病毒蛋白酶的序列,在该序列的切割位点上游和下游有或没有一些或一些氨基酸序列。 a)氨基酸序列所在的前体; b)DNA片段重组,扩增的最终产物和表达载体,允许在已知的诱导型启动子的控制下研究编码逆转录病毒蛋白酶的序列通过将载体和DNA片段与至少一个酵母细胞共转化,c)培养共转化的一个或多个酵母细胞以获得足够数量的转化子以进行敏感性或抗性测试,并回收从co产生的转化子-转化的细胞,在任何合适的培养基上,d)在待测分子的存在下孵育转化体,e)定性或定量定性分析活细胞,和f)推导敏感性或抗性表型。

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