RESUMEN

The C-terminus of the HIV-1 reverse transcriptase heterodimer was reconstructed into a single polypeptide. The construct encodes the p51 thumb (T) and connection (C) subdomains joined through a linker region to the p66 connection (C) and RNase H (R) domain. The TCCR protein was purified from insoluble fractions of Escherichia coli lysates. The TCCR construct maintains Mn(2+)-dependent RNase H activity and specifically cleaves the substrate mimicking the tRNA removal required for second-strand transfer reactions.

Asunto(s)

Transcriptasa Inversa del VIH/química , Ribonucleasa H/química , Western Blotting , Cromatografía en Gel , Electroforesis en Gel de Poliacrilamida , Escherichia coli/metabolismo , Manganeso/química , Modelos Moleculares , Péptidos/química , Unión Proteica , Ingeniería de Proteínas/métodos , Estructura Terciaria de Proteína , ARN de Transferencia/química , Sefarosa/química , Programas Informáticos , Urea/química