Cloning of the full-length rat Nor 1 cDNA using T_4 DNA-ligase-mediated 5' RACE
- VernacularTitle:T_4 DNA连接酶介导的5’RACE法克隆大鼠Nor 1全长cDNA
- Author:
Wei YIN
;
Yijun HUANG
;
Xingwen SU
;
Lingzhi ZHAO
;
Pengxin QIU
;
Guangmei YAN
- Publication Type:Journal Article
- Keywords:
T_4 DNA ligase;
Rapid amplification of cDNA ends;
Nor 1;
Celebellum;
Neurons
- From:
Chinese Journal of Pathophysiology
1986;0(03):-
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To clone the full-length of 75A EST. METHODS: After the extraction of total RNA from primary cultured rat cerebellar granule cell of 7DIV in the medium containing 25 mmol/L KCl, T_4 DNA ligase-mediated 5' RACE was used to retrieve 5' unknown sequence of 75A EST, and the first round 5' RACE PCR product was subcloned into pGEM-T easy vector for sequence and homogeneous analysis. RESULTS: The first round of 5' RACE produce a 2.5 kb band, and 75A EST was identified to be partial sequence of Neuron-derived orphan receptor (Nor1) gene. After two more rounds RACE, we firstly cloned the full-length of Nor-1 cDNA. CONCLUSION: T_4 DNA ligase mediated 5' RACE is an efficient method to retrieve information about the 5' termini of mRNAs, and lay a foundation for further study which role Nor1 play in the cerebellar granule cell differentiation or survive.
