Generated: April 29, 2017
|Title:||Microbial expression of a gene for human growth hormone|
|Abstract:||Described are methods and means for the construction and microbial expression of quasi-synthetic genes arising from the combination of organic synthesis and enzymatic reverse transcription from messenger RNA sequences incomplete from the standpoint of the desired protein product. Preferred products of expression lack bio-inactivating leader sequences common in eukaryotic expression products but problematic with regard to microbial cleavage to yield bioactive material. Illustrative is a preferred embodiment in which a gene coding for human growth hormone (useful in, e.g., treatment of hypopituitary dwarfism) is constructed and expressed.|
|Inventor(s):||Goeddel; David V. (Burlingame, CA), Heyneker; Herbert L. (Burlingame, CA)|
|Assignee:||Genentech Inc. (South San Francisco, CA)|
|Filing Date:||Mar 09, 1982|
|Claims:||1. A method of producing human growth hormone which comprises: |
(a) disposing a culture of bacterial transformants comprising plasmids which, in a transformant bacterium, will express a gene for human growth hormone unaccompanied by the leader sequence of human growth hormone or other extraneous protein bound thereto within a fermenter vessel comprising aeration and agitation means in an aqueous, nutriment-containing fermentation broth;
(b) growing up the culture under aeration and agitation while supplying additional nutriments as required to maintain vigorous growth;
(c) separating the resulting cellular mass from the fermentation broth;
(d) lysing the cells to free the contents thereof;
(e) separating cellular debris from supernatant; and
(f) isolating and purifying human growth hormone contained in the supernatant.
2. A method for producing human growth hormone which method comprises culturing bacterial transformants containing recombinant plasmids which will, in a transformant bacterium, express a gene for human growth hormone unaccompanied by the leader sequence of human growth hormone or other extraneous protein bound thereto, and isolating and purifying said expressed human growth hormone.
3. The method of claim 1 wherein the bacteria are transformant E. coli having a selection characteristic and wherein selection pressure is applied to discourage competition by wild-type E. coli.
Drugs may be covered by multiple patents or regulatory protections. All trademarks and applicant names are the property of their respective owners or licensors. Although great care is taken in the proper and correct provision of this service, thinkBiotech LLC does not accept any responsibility for possible consequences of errors or omissions in the provided data. The data presented herein is for information purposes only. There is no warranty that the data contained herein is error free. thinkBiotech performs no independent verifification of facts as provided by public sources nor are attempts made to provide legal or investing advice. Any reliance on data provided herein is done solely at the discretion of the user. Users of this service are advised to seek professional advice and independent confirmation before considering acting on any of the provided information. thinkBiotech LLC reserves the right to amend, extend or withdraw any part or all of the offered service without notice.