目的:追踪检测转基因防龋可食番茄中外源蛋白的表达稳定性。方法:田间种植含有编码变异链球菌表面蛋白粘附功能A区(Cell surface protein antigen A, PAcA)、表面蛋白免疫活性P区(Cell surface protein antigen P, PAcP)的外源基因pacA-ctxB、pacP-ctxB的转基因番茄种子,获得连续的四代番茄植株F0,F1,F2,F3。在植株的开花坐果期,每七天提取样本,通过生物分子技术检测外源蛋白的表达。结果:1) 四代转基因番茄中含嵌合基因pacA-ctxB的植株中有31株表达外源蛋白;含pacP-ctxB的植株中有28株表达。2) 果实中外源蛋白PAcA/CTB占总蛋白的含量是:F0代0.248%,F1代0.088%,F2代0.103%,F3代0.078%;PAcP/CTB的含量是:F0代0.308%,F1代0.300%,F2代0.278%,F3代0.176%。3) 在开花坐果期的第6~8周,果实中外源蛋白PAcA/CTB含量分别为0.236%,0.276%,0.257%;PAcP/CTB为0.568%,0.600%,0.554%。结论:防龋疫苗中外源蛋白能够较稳定地表达,表达含量较高;PAcP/CTB的表达阳性率及含量均较PAcA/CTB高;外源蛋白在果实中的表达较叶片高;外源蛋白在坐果期的6~8周的表达最高;同株的不同果实同期表达含量及比重有差异。
Objective: To trace the expression stability of exogenous protein PAcA/CTB, PAcP/CTB in the filial generations of the edible anti-caries tomato vaccine. Methods: Four generations of transgenic to-matoes carrying fusion gene pacA-ctxB or pacP-ctxB were planted in the field. Soluble protein of leaves and fruits was extracted every 7 days during fruit-bearing period. Western Blot, BCA and ELISA are used to analyze the content changes of exogenous protein. Results: 1) Exogenous protein is confirmed to be expressed in four generations. 2) The percentage of PAcA/CTB is 0.248% in the F0, 0.088% in the F1, 0.103% in the F2 and 0.078% in the F3. The percentage of PAcP/CTB is 0.308% in the F0, 0.300% in the F1, 0.278% in the F2 and 0.176% in the F3. 3) The percentage of PAcA/CTB during 6th - 8th week was 0.236%, 0.276% and 0.257%; that of PAcP/CTB was 0.568%, 0.600% and 0.554%. Conclusions: Exogenous protein PAcA/CTB and PAcP/CTB were successfully expressed in four filial generations. The proportion of PAcA/CTB was lower than PAcP/CTB. The proportion of PAcA/CTB and PAcP/CTB reached the peak in 6th - 8th week. Expression of exogenous protein in different fruits of one plant at the same time is of significant difference.
目的:追踪检测转基因防龋可食番茄中外源蛋白的表达稳定性。方法:田间种植含有编码变异链球菌表面蛋白粘附功能A区(Cell surface protein antigen A, PAcA)、表面蛋白免疫活性P区(Cell surface protein antigen P, PAcP)的外源基因pacA-ctxB、pacP-ctxB的转基因番茄种子,获得连续的四代番茄植株F0,F1,F2,F3。在植株的开花坐果期,每七天提取样本,通过生物分子技术检测外源蛋白的表达。结果:1) 四代转基因番茄中含嵌合基因pacA-ctxB的植株中有31株表达外源蛋白;含pacP-ctxB的植株中有28株表达。2) 果实中外源蛋白PAcA/CTB占总蛋白的含量是:F0代0.248%,F1代0.088%,F2代0.103%,F3代0.078%;PAcP/CTB的含量是:F0代0.308%,F1代0.300%,F2代0.278%,F3代0.176%。3) 在开花坐果期的第6~8周,果实中外源蛋白PAcA/CTB含量分别为0.236%,0.276%,0.257%;PAcP/CTB为0.568%,0.600%,0.554%。结论:防龋疫苗中外源蛋白能够较稳定地表达,表达含量较高;PAcP/CTB的表达阳性率及含量均较PAcA/CTB高;外源蛋白在果实中的表达较叶片高;外源蛋白在坐果期的6~8周的表达最高;同株的不同果实同期表达含量及比重有差异。
邹怡然,韩 琪,白朋元,管晓燕,白国辉,田 源,陈 筑,程 磊,刘建国. 防龋用转基因番茄外源蛋白表达稳定性研究 The Study of Expression Stability of Exogenous Gene in Transgenic Anti-Caries Tomatoes[J]. 临床医学进展, 2018, 08(06): 529-539. https://doi.org/10.12677/ACM.2018.86088
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