Analisis Integrasi dan Ekspresi Gen Ketahanan terhadap Geminivirus (AV1) pada Tembakau Transgenik

Abstract

Geminivirus infection in several vegetable crops such as tomato and chilli has caused leaf curl and yellowing disease. This disease has significantly impact on yield losses of vegetable production in Indonesia. Genetic engineering technology gives an oppurtunity to develop transgenic plant resistant to Geminivirus. Geminivirus AV1 gene expressing coat protein can be used to develop virus resistant plant. The objective of this research was to identify the integration and expression of AV1 gene in tobacco transgenic plant for resistance to Geminivirus. Analysis of AV1 gene integration was carried out by using polymerase chain reaction (PCR) technique and AV1 gene specific primer as well as tobacco genomic DNA as template. Meanwhile, expression analysis of AV1 gene in transcriptional level was conducted by isolating total RNA, cDNA sintesis using reverse transcriptase-PCR (RT-PCR) and amplification AV1 gene product with the cDNAs as a template. Result of this research showed that out of 60 tobacco plants generation T1 there are 36 plants containing AV1 gene and 18 plants of them showed resistant to Geminivirus. Analysis of AV1 gene expression on 6 selected plants showed that 4 plants expressed AV1 gene that indicated by PCR product from cDNA template.