Cloning and Sequencing of Vial Integration Site in Human Fibroblasts Immortalized by Simian Virus 40
| Project/Area Number |
02807030
|
|---|
| Research Category |
Grant-in-Aid for General Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Research Field |
General medical chemistry
|
|---|
| Research Institution | Univ. of Occupational and Environmental Health |
|---|
Principal Investigator |
GOTOH Sadao Univ. of Occup. &, Biochem., Assistant Professor Environm. Health, 医学部, 助教授 (50131917)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
KARASAKI Yuji Univ. of Occup. &, Biochem., Researcher Environm. Health, 医学部, 助手 (20140907)
HIRANO Hideyasu Univ. of Occup. &, Biochem., Lecturer Environm. Health, 医学部, 講師 (50040241)
|
|---|
| Project Period (FY) |
1990 – 1991
|
| Project Status |
Completed (Fiscal Year 1991)
|
| Budget Amount *help |
¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1991: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1990: ¥1,000,000 (Direct Cost: ¥1,000,000)
|
|---|
| Keywords | Immortalization of Human Cells / DNA sequence at the SV40 integration site / ヒトセンイ芽細胞 / 不死化 / SV40 / ウィルスゲノムの組み込み / 細胞老化 |
|---|
| Research Abstract |
We have analyzed cellular DNA sequences at the viral genome integration site in a human fibroblast cell line VA13 immortalized by simian virus 40(SV40). The compuer analysis of the junctional cellular DNA sequences did not show any homology to the DNA sequences previously reported. This suggests that immortalization by SV40 was not induced by the destruction of any known oncogene or antioncogene at the integration site. We did not find the precise substantial sequence homology at the junctional site between the cellular DNA and SV40 DNA, indicating that the recombination mechanism involved does not require precise sequence homology and therefore, SV40 genome was probably not integrated by homologous recombination. Short direct and inverted repeats of 5 to 29 nucleotides were found in the junctional cellular and SV40 DNA.
Cellular DNA abutting SV40 DNA was found by the Northern blot analysis to be expressed in diploid human fibroblasts and SV40-transformed cells. The nature of this RNA is now under study.
Furthermore, we investigated the other immortalized human cell line named XP(20S)SV. The analysis of the DNA sequence of the SV40 integration site in this cell line revealed the Hind III repeats. However, we couldn't find any oncogene or antioncogene at the viral integration site. Therefore, our final conclusion is that there is not any specific structure required for the establishment of immortalization at the integration site.
|
Report
(3 results)
Research Products
(3 results)
