Molecular cloning of human cardiac troponin I using polymerase chain reaction

FEBS Letters

Barton, Paul J.R.¹  Vallins, William J.¹  Dabhade, Nina¹  Butler-Browne, Gillian²  Yacoub, Magdi H.¹  Brand, Nigel J.¹ 

[1] Department of Cardiothoracic Surgery, National Heart and Lung Institute, Dovehouse Street, London SW7 6LY, UK;INSERM U262, Clinique Universitaire Baudeloque, 123 Port Royal, 75674 Paris, France

关键词: Polymerase chain reaction;    cDNA;    Troponin I;    Amino acid sequence;    TnI;    troponin I (TnIc;    TnIf and TnIs;    cardiac;    fast skeletal and slow skeletal muscle isoforms;    respectively);    PCR;    polymerase chain reaction;    SDS;    sodium dodecyi sulphate;    BSA;    bovine serum albumin;    bp;    base pairs;   

DOI  :  10.1016/0014-5793(90)81234-F

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown