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布拉克须霉八氢番茄红素脱氢酶基因(carB)在卷枝毛霉中的异源表达

Heterologous expression of the Phycomyces blakesleeanus phytoene dehydrogenase gene (carB) in Mucor circinelloides.

作者信息

Ruiz-Hidalgo M J, Eslava A P, Alvarez M I, Benito E P

机构信息

Area de Genética, Departamento de Microbiología y Genética, Universidad de Salamanca, Plaza de los Doctores de la Reina, s/n. 37007, Salamanca, Spain.

出版信息

Curr Microbiol. 1999 Nov;39(5):259-64. doi: 10.1007/s002849900456.

Abstract

A phytoene dehydrogenase-deficient mutant of Mucor circinelloides accumulating only phytoene was transformed with the gene encoding the corresponding enzyme (carB gene) of Phycomyces blakesleeanus. Carotenoids derived from phytoene were detected in the transformants showing that the P. blakesleeanus carB gene complements the M. circinelloides carB mutation. These newly formed carotenoids accumulated in low quantities, indicating that functional complementation was poor. carB mRNA molecules correctly transcribed were detected in the transformants, but they represented a small proportion of the total population of carB-derived mRNAs, mostly constituted by truncated transcripts and by transcripts longer than the transcript that is functional in Phycomyces. These results showed that the P. blakesleeanus carB gene was expressed in M. circinelloides and suggested that the poor complementation observed was owing, at least in part, to the lack of specificity in the recognition of the transcription initiation and termination signals of the P. blakesleeanus carB gene by the M. circinelloides transcriptional machinery.

摘要

用来自布氏毛霉的编码相应酶(carB基因)的基因转化了仅积累八氢番茄红素的卷枝毛霉八氢番茄红素脱氢酶缺陷型突变体。在转化体中检测到了源自八氢番茄红素的类胡萝卜素,这表明布氏毛霉carB基因弥补了卷枝毛霉carB突变。这些新形成的类胡萝卜素积累量很低,表明功能互补性较差。在转化体中检测到了正确转录的carB mRNA分子,但它们在源自carB的mRNA总数中只占一小部分,大部分由截短的转录本和比在布氏毛霉中起作用的转录本更长的转录本组成。这些结果表明布氏毛霉carB基因在卷枝毛霉中表达,并且表明观察到的较差互补性至少部分是由于卷枝毛霉转录机制对布氏毛霉carB基因转录起始和终止信号识别缺乏特异性所致。

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