Premium
Isozyme studies on the association of the herpes simplex virus type 1 thymidine kinase gene with human chromosomes in somatic cell hybrids
Author(s) -
Kit Saul,
Qavi Hamida,
Hazen Marion,
Dubbs Del Rose
Publication year - 1979
Publication title -
international journal of cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.475
H-Index - 234
eISSN - 1097-0215
pISSN - 0020-7136
DOI - 10.1002/ijc.2910240604
Subject(s) - biology , isozyme , microbiology and biotechnology , thymidine kinase , somatic cell , clone (java method) , chromosome , herpes simplex virus , gene , karyotype , genetics , virus , enzyme , biochemistry
To investigate chromosomal site(s) of integration of the herpes simplex virus type 1 (HSV‐1) thymidine kinase (TK) gene in biochemically transformed [HeLa(BU25)/KOS 8–1] cells, these human cells which had been transformed by ultraviolet light‐irradiated HSV‐1 were fused with TK‐negative mouse LM(TK − ) cells, human‐mouse somatic cell hybrid clones (LH81 clones 1–20) were isolated by HATG‐ouabain selection and their chromosomes and isozymes were analyzed. Electrophoretic and serological analyses showed that all 20 clones expressed type‐specific HSV‐1 TK. Isozyme analyses on 29 gene‐enzyme systems representing 22 human chromosomes revealed that all of the HSV‐1 TK‐positive clones expressed human aminoacylase‐1 (ACY‐1) and esterase D (ESD), which have been mapped to human chromosomes 3 and 13, respectively. Other human isozymes were detected in only one to four clones or in none of the clones. Chromosome analyses showed that: (1) the hybrid clones retained only a few human chromosomes; (2) a marker chromosome, designated M7, consisting of a chromosome 17 translocated to the short arm of chromosome 3, occurred in 36 out of the 41 metaphases examined of LH81‐4 clones 1 to 4 and in 31 out of the 33 metaphases examined of LH81–12 clone 10; (3) a modified M7 chromosome, (M7/m), in which the distal 2/3 of the long arm of M7 was translocated to a small acrocentric mouse chromosome, was the only human chromosome found in metaphases of LH81–13 clone 17; and (4) an intact human chromosome 13 was not present in LH81–12 clone 10 or LH81–13 clone 17 cells. Counterselection with BrdUrd resulted in the isolation of subclones lacking HSV‐1 TK, human ACY‐1 and ESD, and the human marker M7 chromosomes. The experiments indicate that the HSV‐1 TK gene is probably associated in HeLa (BU25)/KOS 8–1 cells with marker chromosome M7, but the possibility is not excluded that the segment of human chromosome 13 which codes for ESD is involved.