稻瘟病抗性基因Pi36原核表達(dá)載體構(gòu)建·表達(dá)與鑒定

王玲, 張向明, 劉新瓊

安徽農(nóng)業(yè)科學(xué), 2010, 38(21): 11059-11060  DOI: 10.3969/j.issn.0517-6611.2010.21.010;      追溯原文......本站官方QQ群：62473826

稻瘟病; 抗性基因; 原核表達(dá)載體; 基因克隆

華南農(nóng)業(yè)大學(xué)資源環(huán)境學(xué)院;中南民族大學(xué)生命科學(xué)學(xué)院生物技術(shù)國家民委重點(diǎn)實(shí)驗(yàn)室

[目的]研究稻瘟病抗性基因Pi36的結(jié)構(gòu)和功能。[方法]利用雙酶切構(gòu)建Pi36基因CC、NBS結(jié)構(gòu)域的原核表達(dá)載體，通過菌落PCR鑒定和測(cè)序驗(yàn)證陽性克隆。然后分別收集不同濃度IPTG誘導(dǎo)，不同溫度30和37℃分別培養(yǎng)的大腸桿菌細(xì)胞提取蛋白，利用SDS-PAGE電泳檢測(cè)目的蛋白的表達(dá)情況。[結(jié)果]當(dāng)IPTG濃度為1mmol/L、30℃培養(yǎng)3h，目的蛋白表達(dá)量最大。[結(jié)論]為進(jìn)一步研究稻瘟病抗性基因Pi36的抗病機(jī)理奠定基礎(chǔ)。

Prokaryotic Expression Vector Construction,Expression and Identification of the Blast Resistance Gene Pi36

WANG Ling et al

Journal of Anhui Agricultural Sciences, 2010, 38(21): 11059-11060

Magnaporthe oryza; Resistance gene; Prokaryotic expression vector; Gene cloning

[Objective] The aim was to further study the structure and function of the gene Pi36,which encodes a coiled-coil nucleotide binding site-leucine rich repeat(CC-NBS-LRR).[Method] Prokaryotic expression vector of the gene CC,NBS domain was constructed,and the positive clones were confirmed by colonies PCR and sequencing.The E.coli cells were cultured and collected to extract protein during different concentration of IPTG under 30 and 37 ℃,respectively,and SDS-PAGE was used to detect expression of target prote...

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  稻瘟病抗性基因 Pi36