| Project/Area Number |
05660216
|
|---|
| Research Category |
Grant-in-Aid for General Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Research Field |
General fisheries
|
|---|
| Research Institution | Tokyo University of Fisheries (1994)
University of Miyazaki (1993) |
|---|
Principal Investigator |
AOKI Takashi Tokyo University of Fisheries, Laboratory of Genetics and Biochemistry, Department of Aquatic Biosciences Professor, 水産学部, 教授 (00051805)
|
|---|
| Project Period (FY) |
1993 – 1994
|
| Project Status |
Completed (Fiscal Year 1994)
|
| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1994: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1993: ¥1,500,000 (Direct Cost: ¥1,500,000)
|
|---|
| Keywords | alpha-globin / beta-globin / carp / gene structure / 5'up-stream region of gene / fish cell lines / activin A / promoter activity / alphaグロビン / プロモーター / 遺伝子発現 / CATアッセイ / ネオマイシン耐性 |
|---|
| Research Abstract |
The effect of activin A on the transcriptional activities of the carp alpha-globin promoter sequence in human leukemia cell line K562 and carp fin cell line GF cells was examined using transient expression systems. The level of promoter actibity of carp alpha-globin promoter sequence in the transfected human leukemia cell line K562 cells increased significantly after treatment with activin A.The promoter activity in transfected nonerythroid fish GF cells was not affected by incubation of the cells with activin A.These results showed that activin A increased the transcriptional activities of carp a-globin gene promoter sequence in K562 cells as well as human globin gene promoter sequences.
Neomycin resistance plasmid pSTneo was transfected into four kind of neomycin sensitive fish cell lines. These transformed cell lines were changed to neomycin resistant. The transfected pSTneo integrated into genome of cell lines. These results indicated the neomycin resistance was useful marker for foreign gene transfer to fish cells.
As a result of DNA sequencing analysis of globin family gene, we indicated that the carp alpha-globin genes closely linked the beta-globin gene. The close linkage of carp alpha- and beta-globin genes was proven Southern blot analysis and PCR amplification. The direction of the transcription of the alpha- and beta-globin genes was the opposite. The length between the alpha- and bx e-globin gene translation site was approximately 800bp and the nucleotides of this region was high in of A and T contents. Carp alpha- and beta-globin genes show unique symmetrical arrangement.
|
