本文選題：金針菇 + 農(nóng)桿菌介導(dǎo)法��； 參考：《食用菌學(xué)報(bào)》2017年03期

【摘要】：將釀膿鏈球菌(Streptococcus pyogenes)Cas9序列(SpCas9序列,Addgene#43802)根據(jù)金針菇(Flammulina velutipes)的密碼子偏好性進(jìn)行優(yōu)化,并構(gòu)建FvCas9雙元表達(dá)載體,利用農(nóng)桿菌介導(dǎo)法轉(zhuǎn)化金針菇單核體Dan3,經(jīng)抗性培養(yǎng)基篩選以及潮霉素和Cas9基因PCR擴(kuò)增驗(yàn)證,結(jié)果顯示Cas9基因已經(jīng)成功轉(zhuǎn)入金針菇菌絲體中,轉(zhuǎn)化率為6.84%。
[Abstract]:Streptococcus pyogenes)Cas9 sequence (SpCas9) and Addgene#43802) were optimized according to the codon preference of Flammulina velutipes. and the FvCas9 expression vector was constructed. Agrobacterium tumefaciens mediated transformation of Flammulina velutipes mononuclear Dan3 was carried out. The results showed that the Cas9 gene had been successfully transferred into Flammulina velutipes mycelia and the conversion rate was 6.84 by screening of resistant medium and PCR amplification of hygromycin and Cas9 genes.
【作者單位】： 上海市農(nóng)業(yè)科學(xué)院食用菌研究所 農(nóng)業(yè)部應(yīng)用真菌資源與利用重點(diǎn)開(kāi)放實(shí)驗(yàn)室 國(guó)家食用菌工程技術(shù)研究中心 上海市農(nóng)業(yè)遺傳育種重點(diǎn)實(shí)驗(yàn)室;
【基金】：上海市市級(jí)農(nóng)口系統(tǒng)青年人才成長(zhǎng)計(jì)劃項(xiàng)目[滬農(nóng)青字(2017)第1-13號(hào)];上海市市級(jí)農(nóng)口系統(tǒng)青年人才成長(zhǎng)計(jì)劃項(xiàng)目[滬農(nóng)青字(2017)第1-11號(hào)] 上海市科技興農(nóng)重點(diǎn)攻關(guān)項(xiàng)目[滬農(nóng)科攻字(2015)第5-1號(hào)]資助
【分類(lèi)號(hào)】：S646.15
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本文編號(hào)：1884967