Molecular cloning and characterization of endogenous SV40 dna from human HBL-100 cells

✍ Scribed by C. Saint-Ruf; P. Nardeux; J. Cebiuan; M. Lacasa; C. Lavialle; R. Cassingena

• Publisher: John Wiley and Sons
• Year: 1989
• Tongue: French
• Weight: 738 KB
• Volume: 44
• Category: Article
• ISSN: 0020-7136
• DOI: 10.1002/ijc.2910440230

✦ Synopsis

The human HBL-100 cell line harbours SV, DNA integrated in tandem at a unique site. The SV,, T-antigen expressed in these cells is defective in a function essential to the replication of the viral genome. The integrated SV,, sequences were molecularly cloned in a bacteriophage, and a subclone (plasmid pSVHBI) containing a complete SV, DNA was isolated. As compared to SV, wild-type strain 776, sequence analysis of pSVHBl early region revealed the presence of several DNA alterations. Among these, a point mutation at position 3199, predicting a change at amino-acid 540 of arginine to isoleucine, was shown by marker rescue to be responsible for the deficiency of T-antigen. This novel mutation further delimits one of the T-antigen domains involved in SV, DNA replication. Transfection experiments demonstrated that the transforming activity of the SV, genome from HBL-I00 cells is still preserved. Moreover, several transformed human cell clones thus obtained could be permanently established in culture.