ศูนย์นวัตกรรมเทคโนโลยีหลังการเก็บเกี่ยว

Differential display analysis of gene expression in lightly processed papaya fruit

Y. Karakurt, and D.J. Huber

Proceedings of 26th International Horticultural Congress. Volume of Abstract . Toronto, Canada, 11-17 August, 2002. Abstract S09-P-203. pp. 271.

2002

บทคัดย่อ

Fruits stored as lightly processed (LP) tissue deteriorate much more rapidly than their intact counterparts. In the case of papaya, for example, LP tissue exhibits optimum storage conditions of 4 to 8 days compared with the 2 to 3 weeks for intact fruit. The objective of this study was to isolate and identify genes possibly involved in the deterioration of LP tissue by means of differential display RTPCR. Ripe papaya fuit (skin color: 60-70% yellow) were processed under sanitized conditions at 5 ^oC, and fruit pieces along with intact fruit were both stored for 12 hours at 5 ^oC. Total RNA was isolated from intact and LP papaya fruti and treated with DNAse l to eliminated genomic DNA contaminatiion. Total RNA (0.2 ?g) was used in a reverse transcription reaction with three different one-base-anchored oligo dT primers. High stringency PCR was performed using eight different 18-mer arbitrary primers in combination with the fluorescent-labeled one-base-anchored oligo dT primers. The PCR fluorescent labeled products were separed on a 6% polyacrylamide sequencing gel and scanned with a fluorescent scanner. Confirmation of mRNA transcripts between intact and LP truit revealed that 12 genes were differentially expressed genes were cloned into PCR-TRAP cloning vector. The cloned genes will be sequenced for comparison with know gene sequences already published in the database.