首页> 外国专利> PROTEIN CONSTRUCTS OF MOLONEY MURINE LEUKEMIA VIRUS REVERSE TRANSCRIPTASE (MMLV-RT)

PROTEIN CONSTRUCTS OF MOLONEY MURINE LEUKEMIA VIRUS REVERSE TRANSCRIPTASE (MMLV-RT)

机译:Moloney鼠白血病病毒逆转录酶(MMLV-RT)的蛋白质构建体

摘要

Described are protein constructs of Moloney murine leukemia virus reverse transcriptase (MMLV-RT). In one embodiment the protein construct comprises a Moloney murine leukemia virus reverse transcriptase (MMLV) mature protein sequence; and an expression enhancement sequence at the N-terminus of said MMLV mature protein sequence. The expression enhancement sequence is devised for providing heterologous expression of the protein construct in bacterial cells in absence of chaperone co-expression and/or to provide high-level of expression of the MMLV. In another embodiment the protein construct further comprises a tag/linker combo at the N-terminus of the expression enhancement sequence for assisting in purification of the protein construct. Also described are isolated nucleic acid molecules, vectors and methods for expressing the protein construct in cells. Advantageously these allow large-scale expression of MMLV-RT from cultures of E. coli cells and simplify protocols for purification of MMLV-RT at high purity.
机译:描述是Moloney鼠白血病病毒逆转录酶(MMLV-RT)的蛋白质构建体。在一个实施方案中,蛋白质构建体包含莫尼鼠白血病病毒逆转录酶(MMLV)成熟蛋白质序列;和所述MMLV成熟蛋白质序列的N-末端的表达增强序列。表达增强序列被设计用于在没有伴侣的共表达的情况下提供细菌细胞中蛋白质构建体的异源表达和/或提供MMLV的高含量表达。在另一个实施方案中,蛋白质构建体进一步包含在表达增强序列的N-末端的标签/接头组合,以辅助蛋白质构建体纯化。还描述了分离的核酸分子,载体和用于在细胞中表达蛋白质构建体的方法。有利地,这些允许来自大肠杆菌细胞的培养物的MMLV-RT的大规模表达,并简化了高纯度纯化MMLV-RT的方案。

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