Transformation of the methylotrophic actinomycete Amycolatopis methanolica with plasmid DNA: stimulatory effect of a pMEA300-encoded gene.
نویسندگان
چکیده
Amycolatopsis methanolica contains a 13.29-kb plasmid (pMEA300) present both in the free state and integrated at a unique genomic location. A pMEA300-free derivative (strain WV1) was selected, allowing further analysis of pMEA300-encoded functions. Whole cells of strain WV1 could be transformed at high frequencies (approximately 10(6) transformants per microgram of plasmid DNA) with both circular and linear plasmid DNA, provided that the pMEA300-encoded stf (stimulation of transformation frequency) gene was present. stf would encode a putative protein of 373 amino acids with M(r) 40,201, resembling putative regulatory proteins involved in sporulation of Streptomyces griseus and Streptomyces coelicolor.
منابع مشابه
Identification and functional analysis of the transfer region of plasmid pMEA300 of the methylotrophic actinomycete Amycolatopsis methanolica.
Amycolatopsis methanolica contains a 13.3-kb plasmid (pMEA300) that is present either as an integrated element or as an autonomously replicating plasmid. Conjugational transfer of pMEA300 results in pock formation, zones of growth inhibition that become apparent when plasmid-carrying donor cells develop in a confluent lawn of plasmid-lacking recipient cells. A 6.2-kb pMEA300 DNA region specifyi...
متن کاملA plasmid from the methylotrophic actinomycete Amycolatopsis methanolica capable of site-specific integration.
Amycolatopsis methanolica contains a 13.3-kb plasmid (pMEA300) which is present both in the free state and integrated at a unique genomic location. A 2.1-kb pMEA300 DNA fragment was sequenced, revealing the putative attP site and two open reading frames, xis and int, showing similarity to genes encoding excisionases and integrases, respectively.
متن کاملMolecular cloning with a pMEA300-derived shuttle vector and characterization of the Amycolatopsis methanolica prephenate dehydratase gene.
An efficient restriction barrier for methylated DNA in the actinomycete Amycolatopsis methanolica could be avoided by using a nonmethylating Escherichia coli strain for DNA isolations. The A. methanolica prephenate dehydratase gene was cloned from a gene bank in a pMEA300-derived shuttle vector in E. coli and characterized.
متن کاملMutational analysis of primary alcohol metabolism in the methylotrophic actinomycete Amycolatopsis methanolica
Mutants of the methylotrophic actinomycete Amycolaropsis methanolica unable to grow on methanol as carbon source were isolated and characterized. Mutants specifically affected in methanol utilization were deficient in formaldehyde assimilation. Mutants blocked in the first step of primary alcohol oxidation (Cl-C4) had lost activity of the tetrazolium-dependent alcohol dehydrogenase, a three-com...
متن کاملAmycolatopsis methanolica sp. nov., a facultatively methylotrophic actinomycete.
The generic position of a gram-positive, facultatively methylotrophic actinomycete known as Nocardia sp. strain 239 was determined by comparing reverse transcriptase sequences of 16S rRNA. The assignment of the organism to the genus Amycolatopsis was strongly supported by chemotaxonomic and morphological data. A comparison with the type strains of validly described Amycolatopsis species showed ...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Plasmid
دوره 34 2 شماره
صفحات -
تاریخ انتشار 1995