稻瘟病抗性基因Pi36酵母表達(dá)載體的構(gòu)建與表達(dá)

王玲, 張丹, 張向明, 劉新瓊

湖北農(nóng)業(yè)科學(xué), 2010, 49(7): 1544-1546     追溯原文......本站官方QQ群：62473826

稻瘟病; 抗性基因Pi36; 酵母表達(dá)載體; 構(gòu)建與表達(dá)

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(guó)家民委重點(diǎn)實(shí)驗(yàn)室

稻瘟病抗性基因Pi36編碼卷曲螺旋核苷酸結(jié)合位點(diǎn)-富亮氨酸重復(fù)序列(CC-NBS-LRR)結(jié)構(gòu)的抗病蛋白。為了深入研究該基因的結(jié)構(gòu)和功能，試驗(yàn)利用雙酶切構(gòu)建了Pi36基因CC結(jié)構(gòu)域的酵母表達(dá)載體，通過(guò)菌落PCR鑒定和測(cè)序驗(yàn)證陽(yáng)性克隆。然后分別收集培養(yǎng)至不同OD值的酵母細(xì)胞提取蛋白，利用Western blotting檢測(cè)目的蛋白的表達(dá)情況。結(jié)果表明，目的蛋白成功表達(dá)，并且在OD值為1.0的酵母細(xì)胞中表達(dá)量最大。

Yeast Expression Vector Construction and Expression of the Blast Resistance Gene Pi36

WANG Ling, ZHANG Dan, ZHANG Xiang-ming, LIU Xin-qiong

Hubei Agricultural Sciences, 2010, 49(7): 1544-1546

Magnaporthe oryzae; resistance gene Pi36; yeast expression vector; construction and expression

The blast resistance gene Pi36 encodes a coiled-coil nucleotide binding site-leucine rich repeat (CC-NBS-LRR).To further study its structure and function, yeast expression vector of the gene CC domain was constructed.The positive clones were confirmed by colonies PCR and sequencing.The target protein was extracted from yeast cells with different OD value, and detected by Western blotting.The results showed that this protein expressed successfully, and the highest expression was observed in those yeast cells...

►基因列表◄
  稻瘟病抗性基因 Pi36