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Abstrak :
Telah dilakukan penelitian yang bertujuan untuk melakukan transformasi gen OsERA1 ke kalus padi cv. Taipei 309 menggunakan Agrobacterium tumefaciens. Gen OsERA1 adalah gen yang berperan dalam meningkatkan sensitifitas dari sel penjaga pada stomata terhadap asam absisat. Transformasi gen OsERA1 dilakukan menggunakan Agrobacterium tumefaciens strain LBA4404 yang membawa plasmid rekombinan pCAMBIA1301-OsERA1. Gen OsERA1 telah dikloning sebelumnya pada vektor pengklonaan pGEM-T Easy. Gen dipotong dengan enzim restriksi BamHI dan SalI. Gen diligasikan dengan pCAMBIA 1301 dan ditransformasikan ke dalam Escherichia coli DH5α dihasilkan vektor rekombinan pCAMBIA-ERA1. Plasmid vektor rekombinan pCAMBIA-ERA1 belum berhasil ditransformasi ke dalam Agrobacterium tumefaciens dengan elektroporasi tetapi berhasil dilakukan dengan particle bombardment. ......Research about transformation of OsERA1 gene into rice calli cv. Taipei 309 using Agrobacterium tumefaciens had been done. OsERA1 gene is a gene that has response to enhance sensitivity of guard cell to absicid acid. Transfer of OsERA1gene into calli was carried out using Agrobacterium tumefaciens strain LBA4404 harboring recombinant plasmid pCAMBIA1301-OsERA1. OsERA1 gene had been cloned previously in the pGEM-T Easy cloning vector and for inserting into pCAMBIA1301, the gene was digested from cloning vector using restriction enzymes BamHI and SalI. Furthermore, the gene was ligated into vector pCAMBIA 1301 and transformed into Escherichia coli DH5α in order to obtain recombinant plasmid pCAMBIA-OsERA1. The recombinant plasmid pCAMBIAOsERA1has not sucessfully transformed with electroporation into Agrobacterium tumefaciens yet, but it can be transformed by particle bombardment method.

Abstrak :
Telah dilakukan penelitian yang bertujuan untuk melakukan transformasi gen Osdep1-Tc ke kalus padi cv. Taipei 309 menggunakan Agrobacterium tumefaciens. Transformasi gen Osdep1-Tc dilakukan menggunakan A. tumefaciens strain LBA4404 yang membawa plasmid rekombinan pCAMBIA1301-Osdep1-Tc, mengandung gen reporter (gus), gen nptII dan hptI. Gen Osdep1-Tc yang telah dikloning ke vektor pengklonaan pGEM-T Easy pada penelitian sebelumnya digunakan sebagai sampel untuk kemudian isubkloning ke pCAMBIA 1301 dan ditransformasikan ke dalam Escherichia coli DH5α sehingga dihasilkan vektor rekombinan pCAMBIA-Osdep1-Tc. Vektor rekombinan kemudian dielektroporasi ke A. tumefaciens dan ditransformasi ke kalus embriogenik padi. Aktivitas GUS pada kalus berhasil dideteksi 3 hari setelah infeksi dengan A. tumefaciens. Analisis PCR kalus transforman menunjukkan bahwa gen hptI berhasil terintegrasi dengan stabil pada kelima kalus uji. ......Research about transformation of Osdep1-Tc gene into rice calli cv. Taipei 309 using Agrobacterium tumefaciens had been done. Transformation of Osdep1-Tc was carried out using A. tumefaciens strain LBA4404, harbored recombinant plasmid pCAMBIA1301-Osdep1-Tc, which contained a reporter gene (gus), hptI and nptII gene. Osdep1-Tc gene had been cloned previously into the pGEM-T Easy cloning vector. The gene was being subcloned into pCAMBIA 1301 and transformed into Escherichia coli DH5α in order to obtain recombinant vectors pCAMBIA-Osdep1-Tc. Furthermore, the recombinant vectors was electroporated into A. tumefaciens and transformed into rice embryogenic calli. GUS activity in rice calli was detected 3 days after infection with A. tumefaciens. PCR analysis of the transformant calli revealed that all five calli tested showed a succeeded stable integration of hptI gene.