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- 저자 : Chunyun Guan (검색결과 2 건)
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Prokaryotic expression and protein function of Brassica napus PGIP2 and its genetic transformation
Haiyan HuangFu,Chunyun Guan,Furong Jin,Changfa Yin 한국식물생명공학회 2014 Plant biotechnology reports Vol.8 No.2
Sclerotinia rot is a fungal disease caused bySclerotinia sclerotiorum (Lib.) de Bary, which has severelyreduced rapeseed production worldwide. Polygalacturonase-inhibiting proteins (PIGPs) inhibit theactivity of polygalacturonases, which are secreted duringfungal infection in plants. This study investigated thefunction of the polygalacturonase-inhibitor gene 2 (PGIP2)in sclerotinia rot resistance. The PGIP2 was successfullyexpressed in a prokaryotic system, and recombinant PGIP2protein, purified after enterokinase treatment to remove tagpeptide, inhibited S. sclerotiorum PG activity in vitro. PGIP2 was overexpressed in the susceptible Brassica napuscultivar 98c40 via Agrobacterium-mediated transformation. After inoculation with S. sclerotiorum mycelia, thetransgenic rapeseed demonstrated greatly reduced leafdamage, as compared with their non-transgenic plants. Therefore, the PGIP2 encodes a functional protein andwould be a candidate gene for enhancing Sclerotinia rotresistance.
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Fang Liu,Guiluan Wang,Ruiyang Liu,Chunyun Guan 한국식물생명공학회 2016 Plant biotechnology reports Vol.10 No.6
The gene fatty acid desaturase 2 (FAD2) exists in multiple copies in the Brassica napus genome and encodes an enzyme that catalyzes the conversion of oleic acid to linoleic acid. In the present study, we characterized the regulatory region controlling the expression of an FAD2 gene located on chromosome C5 of Brassica napus and named it BnFAD2-C5. A long intron was found within the 50-untranslated region (50-UTR) of the BnFAD2-C5 gene. This intron, compared with an intron-less control, conferred up to a sixfold increase in green fluorescent protein (GFP) expression in transgenic Arabidopsis, thus suggesting that it makes function through intron-mediated enhancement. The sequence containing the promoter and intron was identified to promote high levels of gene expression in genital organs, particularly in seeds, using qRT-PCR and transgenic Arabidopsis. We identified the different promoter regions responsible for the tissuespecific gene expression through a deletion analysis of the BnFAD2-C5 promoter and a b-glucuronidase and GFP reporter system. The results showed that the -1020 to -319 bp region primarily controls BnFAD2-C5 gene expression in the root, whereas the -1020 to -581 bp region controls expression in the stem, the -581 to -319 bp region controls expression in the leaf, and the -1257 to -1020 bp region probably controls expression in the floral parts. The -319 to -1 bp region is also important, conferring high-level transcription in the seeds. The transcription of BnFAD2-C5 could be induced by salicylic acid and jasmonic acid, and the relative response elements were identified in the -1257 to -1020 bp region and -319 to -1 bp region, respectively.
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