1993 Fiscal Year Final Research Report Summary
Molecular cloning of proliferation-related immediately early gene, hep-1.
| Project/Area Number |
04670797
|
|---|
| Research Category |
Grant-in-Aid for General Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Research Field |
Digestive surgery
|
|---|
| Research Institution | Dokkyo University School of Medicine |
|---|
Principal Investigator |
OHTAKE Hideki Dokkyo Univ.Sch.Med. Associate Professor, 医学部, 助教授 (00049214)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
WATANABE Kazuto Assistant Professor, 医学部, 助手 (80146167)
HASEGAWA Kaoru Associate Professor, 医学部, 講師 (40049250)
|
|---|
| Project Period (FY) |
1992 – 1993
|
| Keywords | Immediate early genes / hep-1 / Rat regenerating liver / Rat hepatocytes in primary culture / PC12 cells / Chromsome mapping / Northern blot hybridization / in situ hybridization |
|---|
| Research Abstract |
Hep-1 cDNA was isolated first from rat hepatocytes lambda gt22A cDNA library by differential plaque hybridization. The size of the cloned cDNA was 1.6Kb. The size of mRNA expressed in rat hepatocytes in primary culture was estimated to be about 2.7Kb by Northern blot hybridization. The gene was expressed in spleen, heart and placenta. Expression in skeletal muscle was weak. In liver or kidney, the gene expression were not observed. Partial sequence analyzes revealed that rat hep-1 (rnhep-1) is found to be the rat PC3 gene isolated from PC12 cells which were stimulated with NGF (Bradbury et al., 1991).
We tried to isolate the human gene corresponding to rnhep-1 by screening the human placent cDNA library with the rnhep-1. The size of mRNA expressed in human placenta cells or human hepatoma cells Hep 3B, was estimated to be bout 3.1Kb by Northern blot hybridization. By plaque hybridization under the mild stringency conditions, 6 positive clones (2.7-3.1Kb) were isolated. Partial nucleotide sequence analyzes revealed that they all coded for the same protein. Comparison of full length cDNAs (3.1Kb) of human gene and rnhep-1 showed 80% identity at the nucleotide level. Hshep-1, the human homologue of rnhep-1, was mapped onto the chromosome 1 by dot blot hybridization. Hshep-1 was expressed in heart, spleen, placenta and endometrium at the proliferating stage of the endometrial cycle. The expression in the endometrium was further analyzed by in situ hybridization. Hshep-1 was expressed in epithelial cells in the uterine glands and those of endometrium.
|
