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Molecular cloning of human cardiac troponin I using polymerase chain reaction
Author(s) -
Vallins William J.,
Brand Nigel J.,
Dabhade Nina,
Butler-Browne Gillian,
Yacoub Magdi H.,
Barton Paul J.R.
Publication year - 1990
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(90)81234-f
Subject(s) - complementary dna , cloning (programming) , molecular cloning , microbiology and biotechnology , polymerase chain reaction , troponin complex , oligonucleotide , rapid amplification of cdna ends , peptide sequence , biology , nucleic acid sequence , gene isoform , troponin i , gene , inverse polymerase chain reaction , biochemistry , nested polymerase chain reaction , medicine , psychiatry , myocardial infarction , computer science , programming language
We have used the polymerase chain reaction (PCR) to synthesise a cDNA encoding part of human cardiac troponin I. Amplification was achieved using fully degenerate sets of oligonucleotides corresponding to conserved regions of amino acid sequence identified in other troponin I isoforms. The cloned PCR fragment was subsequently used to isolate full‐length cDNAs from a cardiac cDNA library. We describe the approach, as a general cloning strategy starting from limited amino‐acid sequence data and report the cloning, and complete amino acid sequence of human cardiac troponin I. Analysis of human development using these clones demonstrates early expression of this gene in the heart.