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Construction and identification of brain derived neurotrophic factor gene cloned into a T-vector in rat

Objective:To construct a T-vector cloned with a BDNF gene of a SD rat.Method:The total RNA of the brain of a SD rat were extracted,the first strand cDNA was synthesized using M-MLV reverse transcriptase and oligo-dT primers,and then BDNF cDNA was amplified from the total cDNA via a PCR using an upstream sense primer and downstream antisense primer,finally the PCR product was subcloned into the pMD18-T simple vector using a metheod of A-T cloning and this complex was prepared for sequencing and other gene clone purpose.Result:A T-vector cloned with the interest BDNF gene was successfully constructed and confirmed by sequencing.Conclusion:The method of cloning BDNF gene with a pMD18-T simple vector is a successful and fast shortcut.