Detection of potyviruses by non-radioactive nucreic acid probe.
| Project/Area Number |
02660043
|
|---|
| Research Category |
Grant-in-Aid for General Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Research Field |
植物保護
|
|---|
| Research Institution | Utsunomiya University |
|---|
Principal Investigator |
NATSUAKI Tomohide (1991) Utsunomiya Univ., Fac. Agric., Associate Professor, 農学部, 助教授 (10134264)
奥田 誠一 (1990) 宇都宮大学, 農学部, 助教授 (90091941)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
OKUDA Seiichi Utsunomiya Univ., Fac. Agric., Associate Professor., 農学部, 助教授 (90091941)
|
|---|
| Project Period (FY) |
1990 – 1991
|
| Project Status |
Completed (Fiscal Year 1991)
|
| Budget Amount *help |
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1991: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1990: ¥1,600,000 (Direct Cost: ¥1,600,000)
|
|---|
| Keywords | non-radioactive probe / Potyviruses / clover yellow vein virus / turnip mosaic virus / distinction of strains / ウエスタンブロット法 / ウイルスRNAーcDNAクロ-ニング / ウイルス系統分類 / フォトビオチン標識プロ-ブ |
|---|
| Research Abstract |
Potyvirus group is one of the biggest group in plant viruses and very important in agriculture. The purpose of this research is to make a specific nonradioactive nucleic acid probe for detecting each polyvirus and its strains. A virus, designated NFU, isolated from bean plants(Phaseolus vulgaris)with severe vine necrosis(Tsurugare-byo)in Fukushima Prefecture was compared with a seed-borne isolate of bean yellow mosaic virus(BYMV-SB)from broad bean. Based on the results of inoculation test, serological test and western blotting, NFU was identified as a strain of clover yellow vein virus(CYVV-NFU). CYVV-RNA was extracted from purified virus preparations and complementary DNA was prepared by the procedure of Gubler and Hoffman. The CDNA was ligated into Smal site of PUC19 plasmid. The E. coli transformed by plasmids containing CDNA inserts were immunoscreened with CYVV antiserum. Three immunopositive clones were obtained. In hybridization test using CDNA probe of CYVV-RNA labeled with photobiotin, a non-radioactive chemical, CYVV was detected very easily and sensitively. Two strains of turnip mosaic virus(TUMV)were also studied. Radish and cabbage strains were distinguished by host range but not by serological method. The CDNA clones were obtained and used as non-radioactive probes.
|
Report
(3 results)
Research Products
(12 results)
