AI Article Synopsis
- Researchers created double-stranded cDNA from poly(A)-containing RNA sourced from bovine anterior pituitary and inserted it into a plasmid for cloning in E. coli.
- Clones that had sequences matching prolactin mRNA were identified using colony hybridization with prolactin cDNA.
- A significant 250 base pair sequence was analyzed, revealing that it encoded parts of the bovine prolactin protein and was used to purify prolactin mRNA from other mRNAs.
Article Abstract
Poly(A)-containing RNA from the bovine anterior pituitary has been used as a template for the enzymatic synthesis of double-stranded cDNA. The resulting double-stranded cDNA was inserted into the Pst I site of pBR322 with the oligo(dG)-oligo(dC) tailing technique and subsequently cloned in E. coli chi 1776. Clones containing sequences complementary to prolactin mRNA were identified by colony hybridization with partially purified prolactin cDNA. A 250 base pair sequence from one prolactin positive clone was extensively characterized and shown to contain the coding information for amino acids 119-192 of authentic bovine prolactin. The recombinant DNA from this clone was covalently attached to diazotized aminocellulose and used to purify prolactin mRNA from a mixture of mRNAs.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC324017 | PMC |
| http://dx.doi.org/10.1093/nar/8.7.1561 | DOI Listing |
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