Isolasi dan Kloning Gen Sitokinin Oksidase (OsCKX) Pengendali Sifat Produktivitas dari Padi Varietas Ciherang

Abstract

Gn1a/OsCKX gene (grain number 1a/Cytokinin Oxidase) of rice (Oryza sativa L.) that encodes cytokinin oxidase has been identified as a quantitative trait loci that contribute to an increase in the number of grains of rice. This study aimed to isolate an OsCKX gene from rice cv. Ciherang. Amplification of OsCKX gene was done by PCR (Polymerase Chain Reaction) technique using specific primer pairs and cDNA template of rice cv. Ciherang. Subsequently, OsCKX gene fragment was cloned into a cloning vector pGEM-T Easy using T4-ligase enzyme and transformed into competent cell Escherichia coli Dh5α through heat shock method, and grown on LB agar medium containing ampicillin. Result showed that amplification with specific primers of OsCKX gene generated an amplicon fragment with size of ± 2000 bp. The gene fragment was succesfully cloned into pGEM-T vector and transformed into cells of E. coli Dh5α. Verification of recombinant clone by cutting using two restriction enzymes SmaI and KpnI regenerated two bands of 3015 bp (pGEM-T Easy vector) and 2000 bp (OsCKX gene fragment).