Abstract
The progesterone receptor is a type of protein present inside cells, which is activated by the hormone
progesterone (type of steroid hormone). In humans, a single PGR gene found on chromosome 11q22 encodes
progesterone. There are two main types, A and B the difference lies in their molecular weight. Progesterone
receptor isoform B functions tissue-specifically to mediate reproductive functions of progesterone in the present
study we did cloning of progesterone receptor isoform B in expression vector CMV-eGFP. Gene cloning provides
information about nucleotide sequence. Other motive of this is to manipulate the gene in various ways like
genetic transformation. Genomic DNA was isolated from human blood. For cloning of PR-B and CMV-eGFP,
restriction and ligation were done using appropriate conditions and concentrations. The resulting DNA was
transformed into host organism (E.coli, DH5α). These were grown on kanamycin resistant agar plated. Colonies
containing ligated PR-B with plasmid were then screened, purified and analysed using standard protocol. The
desired CMV-eGFPC1-PR-B construct of 7523bp was obtained and stored at -20oC.
SAQIBA KHALID , NAILA MALKANI. (2016) Isolation and cloning of human progesterone receptor isoform-b in CMV-eGFP vector, Biologia – Journal of Biological Society of Pakistan, Volume 62 (I), Issue 1.
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