本文選題：變異鏈球菌　切入點：霍亂毒素B亞單位　出處：《上�？谇会t(yī)學(xué)》2014年03期 　論文類型：期刊論文

【摘要】：目的:在獲得含編碼變異鏈球菌表面蛋白PAcP和霍亂毒素B亞單位融合基因的轉(zhuǎn)基因番茄原代種子的基礎(chǔ)上,應(yīng)用分子生物學(xué)技術(shù)檢測外源基因在轉(zhuǎn)基因植株中的表達(dá),測定目的蛋白的含量。方法:PCR篩選含編碼變異鏈球菌表面蛋白PAcP與霍亂毒素B亞單位融合基因的轉(zhuǎn)基因番茄植株;提取番茄果實總蛋白,用BCA試劑盒測定番茄果實總蛋白含量;通過Western印跡檢測外源蛋白的表達(dá)情況,并用ELISA法對外源目的蛋白含量進(jìn)行測定。結(jié)果:PCR擴(kuò)增分析可見1.6 kb特異性擴(kuò)增條帶,出現(xiàn)特異性條帶的植株占總檢測植株的55.6%;轉(zhuǎn)基因番茄總蛋白含量為3.93 mg/mL,Western印跡結(jié)果顯示,在PVDF膜上,約60 kD處出現(xiàn)特異性條帶;ELISA測得表達(dá)的目的蛋白占番茄可溶性總蛋白的0.18%。結(jié)論:含編碼變異鏈球菌表面蛋白PAcP和霍亂毒素B亞單位融合基因的轉(zhuǎn)基因番茄子代植株能有效表達(dá)外源蛋白。
[Abstract]:Objective: on the basis of obtaining transgenic tomato primary seeds containing fusion gene of Streptococcus mutans surface protein PAcP and cholera toxin B subunit, the expression of exogenous genes in transgenic plants was detected by molecular biology technique. Methods the transgenic tomato plants containing fusion gene encoding Streptococcus mutans surface protein (PAcP) and cholera toxin B subunit (CTB) were selected and the total protein of tomato fruit was extracted and the total protein content of tomato fruit was determined by BCA kit. The expression of exogenous protein was detected by Western blotting, and the content of exogenous protein was determined by ELISA. The plants with specific bands accounted for 55.6% of the total tested plants, and the total protein content of transgenic tomato was 3.93 mg / mL Western blot. The target protein expressed by Elisa was 0.18% of the total soluble protein in tomato. Conclusion: transgenic tomato progenies containing fusion gene encoding Streptococcus mutans surface protein PAcP and cholera toxin B subunit are obtained by Elisa. The strain can express foreign protein effectively.
【作者單位】： 遵義醫(yī)學(xué)院口腔學(xué)院;貴州省高等學(xué)�？谇患膊⊙芯刻厣攸c實驗室;貴陽市口腔醫(yī)院;
【基金】：國家自然科學(xué)基金(30160086) 貴州省科技支撐計劃項目[黔科合SY字(2012)3086號] 貴州省第六批科技創(chuàng)新人才團(tuán)隊資助項目[黔科合人才團(tuán)隊(2013)4026號] 貴州省科學(xué)技術(shù)基金項目[黔科合J字LKZ(2011)41號]~~
【分類號】：R780.2

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