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肌苷菌核酸代谢关键酶缺失和形成选育腺苷菌的研究 被引量:8
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作者 柏建新 朱晓宏 +4 位作者 张一平 王红连 邓崇亮 杜郭君 罗先勇 《微生物学通报》 CAS CSCD 北大核心 2003年第2期52-56,共5页
以肌苷产生菌BacillusSubtilisJSIM 1 0 1 9为出发菌株 ,根据B subtilis核酸代谢理论设计不同筛选模型 ,用物理、化学诱变剂对亲株进行诱变处理 ,先后有序地获得不同关键酶的缺失或回复即特殊的营养缺陷型以及抗某些代谢类似物的突变株 ... 以肌苷产生菌BacillusSubtilisJSIM 1 0 1 9为出发菌株 ,根据B subtilis核酸代谢理论设计不同筛选模型 ,用物理、化学诱变剂对亲株进行诱变处理 ,先后有序地获得不同关键酶的缺失或回复即特殊的营养缺陷型以及抗某些代谢类似物的突变株 ,解除终产物对代谢物的抑制和阻遏 ,获得了几株黄嘌呤营养缺陷型并对 8氮鸟嘌呤具有抗性的突变株X 1 3等 ,获得的突变株经单菌分离后得到X 1 3 4 ,36℃培养 72h在培养基中最高积累 1 2 4 3g/L腺苷。 展开更多
关键词 肌苷菌 核酸代谢酶 缺失 选育 腺苷菌 黄嘌呤 肌苷酸脱氢 腺苷酸脱氨 乌苷酸还原
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siRNA表达载体对hTERT基因的效应评价 被引量:2
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作者 赵矫 余红 +2 位作者 赵小平 税青林 曾永秋 《陕西医学杂志》 CAS 2009年第1期7-10,共4页
目的:构建人端粒酶逆转录酶(hTERT)基因的RNA干扰表达载体,探讨其对人乳腺癌MCF-7细胞生长及其hTERT基因表达的影响。方法:设计针对hTERT基因的干扰靶序列并构建重组siRNA表达载体pGenesil-hTERT。酶切测序鉴定后,脂质体转染MCF-7细胞,... 目的:构建人端粒酶逆转录酶(hTERT)基因的RNA干扰表达载体,探讨其对人乳腺癌MCF-7细胞生长及其hTERT基因表达的影响。方法:设计针对hTERT基因的干扰靶序列并构建重组siRNA表达载体pGenesil-hTERT。酶切测序鉴定后,脂质体转染MCF-7细胞,应用RT-PCR和Weston Blot法检测hTERT基因的表达情况,应用流式细胞仪测定细胞凋亡率。结果:酶切电泳测序分析表明插入序列正确,重组质粒构建成功,且转染MCF-7细胞后,hTERT mRNA和蛋白质表达水平显著抑制,细胞凋亡率较对照组明显升高(P<0.01)。结论:以DNA质粒为载体的siRNA能有效抑制hTERT基因的表达,进而抑制乳腺癌MCF-7细胞生长、促进细胞凋亡。 展开更多
关键词 乳腺肿瘤/病理生理学 核糖核酸类/代谢 端粒 DNA指导的RNA聚合
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Recent advance in SNP identifying methods and individualized medication 被引量:2
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作者 彭锐 张洪 张英 《Journal of Chinese Pharmaceutical Sciences》 CAS CSCD 2014年第10期731-738,共8页
Polymorphisms associated with genes coding for a variety of drug-metabolizing enzymes (DMEs) and associated transport proteins can influence the drug metabolism rate of individuals, potentially affecting the efficac... Polymorphisms associated with genes coding for a variety of drug-metabolizing enzymes (DMEs) and associated transport proteins can influence the drug metabolism rate of individuals, potentially affecting the efficacy of drug and the occurrence of adverse reactions. Single nucleotide polymorphisms (SNPs) are prevalent in all types of genetic variations. Reliable SNP genotyping provides excellent markers for detecting genetic polymolphisms, genetic disorders, and resistance of pathogen to drug, which are needed for the genetic diagnosis of disease and subtle genetic factors. With a large number of SNP genotyping studies being conducted, a lot of novel SNP identifying methods have been developed. Several SNP genotyping methods and techniques have been introduced for clinical test. These include TaqMan drug metabolism genotyping assays, pH-sensing semiconductor system, high-resolution melting curve analysis (HRM) of polymerase chain reaction (PCR) amplicons, novel multiplexed electrochemical biosensor with non-fouling surface, DNA hybridization detection using less than 10-nm gap silicon nanogap structure, tetra-primer ARMS-PCR method, acoustic detection of DNA conformation in genetic assays combined with PCR, microbeads-mass spectrometry (MEMS)-based approach, and liquid chromatography-electrospray ionization mass spectrometry. Personalized medicine has changed the conventional ways of using drugs according to experiences. It focuses on making the individualized pattern for each individual based on their own characteristics. Lots of researchers are using the analysis of clinical samples to explain the relationship between the drug adverse reactions and genetic polymorphisms. But it takes a long time from collecting the blood samples for DNA extraction and genotyping to getting results on the side effect of drug through clinical study. Therefore, it is desirable to develop improved in vitro methods to study the drug metabolizing-enzymes and transport protein genetic polymorphisms. 展开更多
关键词 Single nucleotide polymorphism GENOTYPING Drug-metabolizing enzymes Transporters PHARMACOGENOMICS
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