The construction of the hammerhead ribozyme genes targeting against apple scar skid viroid and its activity detection in vitro / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 25-29, 2002.
Article
in Chinese
| WPRIM
| ID: wpr-231380
ABSTRACT
The genes of short armed hammerhead ribozyme targeting against two sites on positive strand (194-196) and negative strand (89-91) of ASSVd were designed, synthesized and cloned according to the action manner of hammerhead ribozyme. The full lengths of the genes are 42 bp (RzASSVd(+)) and 40 bp (RzASSVd(-)). After transcription in vitro, the ASSVd positive and negative RNA labeled with 32P were mixed with the ribozyme transcript and incubated 3-4 h at 50 degrees C or 37 degrees C. The results were assayed on 8% PAGE (containing 8 mol/L urea) and autoradiogrammed. As predicted, the transcript of the active RzASSVd(-) could cleave the ASSVd negative strand RNA with a high activity but had no cleavage effect on the ASSVd positive strand. The transcript of the RzASSVd(+) gene could cleave the ASSVd positive strand but its cleavage activity was very low. As the same, it cannot cleave the negative strand either. On the base of the result, we construct dimmer ribozyme gene pGEMRzASSVd(+/-) containing both RzASSVd(+) and RzASSVd(-).
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Plant Diseases
/
Substrate Specificity
/
Transcription, Genetic
/
Viroids
/
Virology
/
RNA, Viral
/
RNA, Catalytic
/
Cloning, Molecular
/
Malus
/
Therapeutic Uses
Type of study:
Diagnostic study
Language:
Chinese
Journal:
Chinese Journal of Biotechnology
Year:
2002
Type:
Article
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