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Cloning of the full-length rat Nor 1 cDNA using T_4 DNA-ligase-mediated 5' RACE / 中国病理生理杂志
Article in Zh | WPRIM | ID: wpr-522305
Responsible library: WPRO
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.
Key words
Full text: 1 Database: WPRIM Aspects: Determinantes_sociais_saude Language: Zh Journal: Chinese Journal of Pathophysiology Year: 1986 Document type: Article
Full text: 1 Database: WPRIM Aspects: Determinantes_sociais_saude Language: Zh Journal: Chinese Journal of Pathophysiology Year: 1986 Document type: Article