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Expression of the canine relaxin precursor in Pichia pastoris host: A canine pregnancy specific protein.

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Title Expression of the canine relaxin precursor in Pichia pastoris host: A canine pregnancy specific protein.
Not Available
 
Creator P Harikrishna
S K Bhure
A M Shende
Praveen Singh
A M Pawde
 
Subject relaxin
canine
 
Description Not Available
In canines, a physical examination and ultrasound technique remain the most common methods of pregnancy diagnosis. The rapid detection of pregnancy is important in canines. There are several reports of peptide hormone relaxin to be exclusively produced in pregnant bitches and not by non-pregnant or pseudo pregnant bitches. We present here the expression of canine relaxin in Pichaia pastoris GS115 host. The cDNA prepared using canine placental total RNA was used to amplify 432 bp relaxin gene, which was subsequently cloned in pPICZ-alphaA vector and followed by transformation into Pichia pastoris GS115 strain by electroporation. The zeocin resistant recombinant clones were confirmed by colony PCR and sequencing. One of the positive clones was selected for protein expression in culture medium. The supernatant was taken and precipitated using 20% TCA in acetone. The recombinant relaxin precursor protein showed a mol wt of 24 kDa, which is higher than the expected 19 kDa and that could be due to glycosylation in the Pichia pastoris. The 24 kDa recombinant protein was detected up to 48 h of post-induction. This is the first report describing the expression of recombinant canine relaxin in Pichia pastoris. The polyclonal antiserum against canine relaxin would be useful for detection of pregnancy in bitches and also in assessing the prognosis of canine mammary tumors
Not Available
 
Date 2023-06-21T04:34:14Z
2023-06-21T04:34:14Z
2017-01-01
 
Type Research Paper
 
Identifier Not Available
Not Available
http://krishi.icar.gov.in/jspui/handle/123456789/78478
 
Language English
 
Relation Not Available;
 
Publisher NIScPR