转基因水稻蛋白试纸条与PCR定性检测技术比较研究

以现有的5个转基因水稻品系汕优63(TT51-1)、科丰6号(KF6)、Ⅱ优科丰6号(Ⅱ优KF6)、克螟稻(KMD)、M12为研究材料,分别采用PCR定性法和蛋白试纸条法2种方法进行检测,比较了2种方法的一致性、灵敏度、准确性,并明确了2种检测方法的适用范围。

PCR结合核酸斑点杂交技术检测转基因番茄研究

基于转基因番茄的安全性评价,建立健全番茄转基因检测体系尤为迫切。文章采用转基因作物检测常用的传统定性PCR技术和核酸斑点杂交技术,针对转基因番茄中常用的35S(花椰菜花叶病毒启动子)、NOS(胭脂碱合成酶终止子),NPTII(新霉素转移酶基因),35S-EFE(35S和番茄EFE基因的交联结构)、以及Lat52(番茄内参基因),检测了阳性对照质粒PBI121、华蕃一号样品和阴性对照合作903番茄。从华蕃一号中成功检出35S、NOS、NPTII、35S-EFE和内参基因Lat52,阴性对照番茄合作903中只有内参基因Lat52检出。试验重复性良好,检测灵敏度高,两种方法相互补充,排除假阳性。核酸斑点杂交一次可检出多个外源基因,增加了检测通量,为我国番茄转基因检测提供参考。

高分子量谷蛋白亚基转基因小麦检测技术的研究

针对该实验材料转入的外源基因选择了标准中通用的外源基因CaMV35S,NOS,bar和植物内源基因tRNALeu作为特异性引物,对影响定性PCR检测(反应体系:氯化镁浓度,引物和模板浓度;反应条件:退火温度和时间,循环数等)的主要条件进行了实验优化和选择,同时对PCR产物进行琼脂糖凝胶电泳分析。结果表明,可以检测到CaMV35S,NOS,bar外源基因预期大小的目的条带,表明定性PCR检测所优化的条件适合转基因小麦。实验还测定了定性PCR最低检测灵敏度为1%(w/w)。同时对转入外源目的基因进行了表达蛋白的检测,蛋白电泳结果表明检测出1Dx5亚基和1Dy10亚基。

Study on the Correlation between the Expression of Angiogenic Factor VEGF and Microsatellite Instability in Gastric Adenocarcinoma

Objective: To investigate the correlation between the microsatellite instability (MSI) and the expression of vascular endothelial growth factor (VEGF) in gastric adenocarcinoma. Methods: PCR SSCP method was used to detect MSI of thirty cases with gastric adenocarcinoma at five loci in each patient. Expression of VEGF was examined by the method of immunohistochemistry. Results: The positive of MSI was in 13 patients out of 30 patients (43.4%) in our study. Positive VEGF Immunostaining was detected in 18 patients (60.0%). VEGF was decreased in microsatellite instability high (MSI H) gastric adenocarcinoma. Conclusion: MSI H and microsatellite stable (MSS) gastric adenocarcinoma may follow a different pathway of angiogenesis. The low frequency of VEGF expression among MSI H cancer may partially explain why these cancers are less aggressive.