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Analysis of integrated hepatitis B virus DNA and cellular flanking sequences cloned from a hepatocellular carcinoma

✍ Scribed by Hisashi Matsumoto; Tetsuo Yoneyama; Keiji Mitamura; Toshiaki Osuga; Hiroto Shimojo; Tatsuo Miyamura

Publisher: John Wiley and Sons
Year: 1988
Tongue: French
Weight: 631 KB
Volume: 42
Category: Article
ISSN: 0020-7136
DOI: 10.1002/ijc.2910420102

No coin nor oath required. For personal study only.

✦ Synopsis

By digestion with Hindlll restriction enzyme, a human hepatocellular carcinoma was shown to contain only 2 hepatitis B virus (HBV) DNA inserts. Both Hindlll fragments (8 and 16 kb) were molecularly cloned and the structures of HBV DNA and adjacent host sequences were analyzed. One clone (AYH 8) contained part of proS(1) through the entire S gene and the other (AYH 16) had the middle section of the S to the end of X gene of HBV DNA. No gross rearrangements were observed in either HBV or cellular DNA sequences in XYH 16 clone. However, the 8 kb Hindlll fragment was considered to be amplified together with flanking cellular sequences. Furthermore, the HBV DNA was inte rated into cellular genome at the Alu repeated sequences in X&H 8.

Nucleotide sequencing

Regions containing virus-host junctions were identified by fine restriction mapping of subcloned DNAs. Restriction fragments (HincII-TaqI and AccI-HincII fragments of XYH 8, AvaII-AvaII and PstI-HincII fragments of XYH 16) were subcloned into single-stranded phage vectors M13mp18 or M13mp19. Nucleotide sequences were determined by the dideoxy chain termination method (Sanger et al., 1977).

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