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filingDate 1981-01-19-04:00^^<http://www.w3.org/2001/XMLSchema#date>
publicationDate 1981-07-30-04:00^^<http://www.w3.org/2001/XMLSchema#date>
publicationNumber GB-2067574-A
titleOfInvention Microbiologically prepared polypeptide with the amino acid sequence of proinsulin of a primate, DNA and plasmids which code for this sequence, microorganisms which contain this genetic information, and processes for their preparation
abstract Human hormones are required to an increasing extent for therapy, and methods involving recombinant DNA have been proposed for the production of certain mammalian hormones. There is provided a microbially prepared polypeptide, all or part of which comprises the amino acid sequence of proinsulin of a primate. There is also provided cDNA which codes for the above polypeptide, prepared using mRNA of a primate, and a plasmid comprising this cDNA, and a microorganism comprising such a plasmid. The polypeptide may be prepared by obtaining mRNA from a primate, especially from the pancreas of a primate, producing double stranded DNA therefrom, incorporating the DNA into a plasmid and the plasmid into microorganisms, and selecting and culturing those microorganisms that produce the polypeptide. In the production of the mRNA, proteolytic digestion of the pancreatic tissue is preferably carried out in the presence of mammalian serum, especially bovine serum, and in caesium chloride centrifugation of a beta -cell lysate, preferably a caesium chloride-free lysate is arranged on an aqueous chloride solution prior to centrifugation. This technique is applicable to the isolation of RNA from any source.
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