Poster: Late and Moved Abstracts

Abs # 1381: PGIP cloning from Brassica campestris (syn. B. rapa perkinensis)

Presenter:	Kim, Goon-Bo , mycorr@empal.com
Authors	Kim, Goon-Bo  (A)   Byung-Ho, Whang  (B)   Jong-Ghi, Kim  (B)   Shinje, Kim  (A)
Affiliations:	(A): FNP Corp. (B): Joong-Ang University

Polygalacturonase inhibiting protein is known to play a key role in the response to fungal infection. It inhibits the action of fungal polygalacturonase, which macerates the cell wall pectin, to not depolymerase cell wall matrix. To clone the PGIP genes in B. campestris, degenerate PCR, cDNA library screening, and inverse PCR technique were used. Degenerate PCR products which was PGIP homolog from B. napus was used to screen the 77K cDNA clones. Among total of 63 positive cDNA clones, long-sized clones were sequenced and assembled to produce CAP3 cDNA contig, and clustered to 5 different ORF sequences and 7 different 3'UTR. As none of full length cDNA was cloned, BAC library of 12X genome size was screened to select 8 clones. Among them 4 clones were used in inverse PCR to amplify the upstream regions of each classes of pgip cDNAs. 5' missing end of each cDNA were amplified by 5'-end RACE. 5 PGIP sequences(termed as BcPGIP1, 2, 3, 4, and 5) were strongly similar to PGIP1, 2, 3, and 4 from B. napus by 60%-99%. As a result, Brassica campestris( syn. B. rapa subs. perkinensis) was revealed to have diverse classes of PGIP.