人bFGF基因轉(zhuǎn)化大豆毛狀根表達(dá)體系的實(shí)驗(yàn)研究
發(fā)布時(shí)間:2018-07-02 20:42
本文選題:人堿性成纖維細(xì)胞生長(zhǎng)因子(hbFGF) + 大豆; 參考:《吉林大學(xué)》2007年博士論文
【摘要】: 本研究利用基因重組技術(shù)在大豆毛狀根中表達(dá)人bFGF。將人bFGF基因克隆到質(zhì)粒pCAMBIA1301載體上,以大豆子葉節(jié)和胚軸為外植體,通過(guò)發(fā)根農(nóng)桿菌C58C1介導(dǎo)轉(zhuǎn)入到大豆中,,經(jīng)潮霉素抗性篩選,獲得潮霉素抗性的毛狀根。PCR法擴(kuò)增檢測(cè)人bFGF基因的整合。提取毛狀根中蛋白,用Western blot印跡分析人bFGF的表達(dá)。陽(yáng)性毛狀根中整合并表達(dá)人bFGF基因。SDS-PAGE顯示樣品相對(duì)分子量約為18kDa,與人bFGF標(biāo)準(zhǔn)品相對(duì)分子量一致,證明人bFGF基因在大豆的毛狀根中正確表達(dá)。對(duì)轉(zhuǎn)化毛狀根規(guī);瘧腋∨囵B(yǎng)條件進(jìn)行了優(yōu)化,確定了最佳的培養(yǎng)條件,為利用大豆毛狀根作為生物反應(yīng)器工業(yè)化生產(chǎn)天然藥物奠定實(shí)驗(yàn)基礎(chǔ)。
[Abstract]:The aim of this study was to express human bFGF in soybean hairy roots by gene recombination technique. The human bFGF gene was cloned into plasmid pCAMBIA1301 and transformed into soybean by Agrobacterium tumefaciens C58C1, and was screened by hygromycin resistance. Hygromycin resistant hairy roots were amplified by PCR to detect the integration of human bFGF gene. The protein was extracted from hairy root and the expression of human bFGF was analyzed by Western blot. The expression of human bFGF gene. SDS-PAGE in the positive hairy root showed that the relative molecular weight of the sample was about 18kDa. it was proved that the human bFGF gene was correctly expressed in the hairy root of soybean. The optimal culture conditions of the transformed hairy roots were optimized and the optimum culture conditions were determined, which laid the experimental foundation for the industrialized production of natural drugs by using soybean hairy roots as bioreactor.
【學(xué)位授予單位】:吉林大學(xué)
【學(xué)位級(jí)別】:博士
【學(xué)位授予年份】:2007
【分類號(hào)】:R346
【引證文獻(xiàn)】
相關(guān)會(huì)議論文 前1條
1 李天航;楊晶;王艷芳;郭詠昕;李昌禹;李海燕;李校X;;發(fā)狀根在藥用植物基因工程研究進(jìn)展[A];2010年中國(guó)藥學(xué)大會(huì)暨第十屆中國(guó)藥師周論文集[C];2010年
相關(guān)碩士學(xué)位論文 前2條
1 李天航;人參發(fā)狀根的誘導(dǎo)培養(yǎng)體系建立及TSPG對(duì)HepG2細(xì)胞抑制作用[D];吉林農(nóng)業(yè)大學(xué);2011年
2 劉蓓蓓;煙草根特異性啟動(dòng)子植物表達(dá)載體的構(gòu)建及其對(duì)紅景天發(fā)根的遺傳轉(zhuǎn)化[D];吉林農(nóng)業(yè)大學(xué);2011年
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