以分离自东北自然发酵大酱中的一株耐盐植物乳杆菌FS5-5(Lactobacillus plantarum FS5-5)为实验对象,在转录水平上对该菌株盐胁迫相关基因表达进行了研究。结果表明:在对数生长期,表达显著下调的共29个基因,分别为参与碳水化合物转运和...以分离自东北自然发酵大酱中的一株耐盐植物乳杆菌FS5-5(Lactobacillus plantarum FS5-5)为实验对象,在转录水平上对该菌株盐胁迫相关基因表达进行了研究。结果表明:在对数生长期,表达显著下调的共29个基因,分别为参与碳水化合物转运和代谢的4个基因,氨基酸转运和代谢的9个基因,维生素代谢的3个基因,核苷酸代谢的6个基因,遗传信息翻译、核糖体结构和形成的7个基因;表达显著上调的有参与碳水化合物转运和代谢的4个基因。这些变化可能与L.plantarum FS5-5的盐胁迫机制密切相关。选取参与维生素代谢的3个基因和核苷酸代谢的1个基因,通过实时定量聚合酶链式反应对转录组学结果进行验证,结果表明两种方法中基因表达趋势一致。实验对L.plantarum FS5-5的盐胁迫反应进行了比较全面的分析,为提高工业生产中菌株的耐受性提供了理论依据。展开更多
AIM:To establish the more feasible and sensitive assessment approach to the detection of adefovir (ADV) resistance-associated hepatitis B virus (HBV) quasispecies.METHODS: Based on the characteristics of rtA181V/T and...AIM:To establish the more feasible and sensitive assessment approach to the detection of adefovir (ADV) resistance-associated hepatitis B virus (HBV) quasispecies.METHODS: Based on the characteristics of rtA181V/T and rtN236T mutations, a new approach based on real-time fluorescent quantitative polymerase chain reaction (RT-PCR) was established for the detection of ADV-resistant HBV quasispecies, total HBV DNA, rtA181 and rtN236 mutations in blood samples from 32 chronic hepatitis B (CHB) patients with unsatisfactory curative effect on ADV and compared with routine HBV DNA sequencing.RESULTS: Both the sensitivity and specificity of this new detection approach to ADV-resistant HBV quasispecies were 100%, which were much higher than those of direct HBV DNA sequencing. The approach was able to detect 0.1% of mutated strains in a total plasmid population. Among the 32 clinical patients, single rtA181 and rtN236T mutation and double rtA181T and rtN236T mutations were detected in 20 and 8, respectively, while ADV-resistant mutations in 6 (including, rtA181V/T mutation alone in 5 patients) and no associated mutations in 26.CONCLUSION: This new approach is more feasible and efficient to detect ADV-resistant mutants of HBV and ADV-resistant mutations before and during ADV treatment with a specificity of 100% and a sensitivity of 100%.展开更多
文摘以分离自东北自然发酵大酱中的一株耐盐植物乳杆菌FS5-5(Lactobacillus plantarum FS5-5)为实验对象,在转录水平上对该菌株盐胁迫相关基因表达进行了研究。结果表明:在对数生长期,表达显著下调的共29个基因,分别为参与碳水化合物转运和代谢的4个基因,氨基酸转运和代谢的9个基因,维生素代谢的3个基因,核苷酸代谢的6个基因,遗传信息翻译、核糖体结构和形成的7个基因;表达显著上调的有参与碳水化合物转运和代谢的4个基因。这些变化可能与L.plantarum FS5-5的盐胁迫机制密切相关。选取参与维生素代谢的3个基因和核苷酸代谢的1个基因,通过实时定量聚合酶链式反应对转录组学结果进行验证,结果表明两种方法中基因表达趋势一致。实验对L.plantarum FS5-5的盐胁迫反应进行了比较全面的分析,为提高工业生产中菌株的耐受性提供了理论依据。
基金Supported by The fund from Health Project of Jiangsu Province,No.H200711the AIDS,Hepatitis B and Other Infectious Diseases Prevention Program,No.2009ZX10004-712
文摘AIM:To establish the more feasible and sensitive assessment approach to the detection of adefovir (ADV) resistance-associated hepatitis B virus (HBV) quasispecies.METHODS: Based on the characteristics of rtA181V/T and rtN236T mutations, a new approach based on real-time fluorescent quantitative polymerase chain reaction (RT-PCR) was established for the detection of ADV-resistant HBV quasispecies, total HBV DNA, rtA181 and rtN236 mutations in blood samples from 32 chronic hepatitis B (CHB) patients with unsatisfactory curative effect on ADV and compared with routine HBV DNA sequencing.RESULTS: Both the sensitivity and specificity of this new detection approach to ADV-resistant HBV quasispecies were 100%, which were much higher than those of direct HBV DNA sequencing. The approach was able to detect 0.1% of mutated strains in a total plasmid population. Among the 32 clinical patients, single rtA181 and rtN236T mutation and double rtA181T and rtN236T mutations were detected in 20 and 8, respectively, while ADV-resistant mutations in 6 (including, rtA181V/T mutation alone in 5 patients) and no associated mutations in 26.CONCLUSION: This new approach is more feasible and efficient to detect ADV-resistant mutants of HBV and ADV-resistant mutations before and during ADV treatment with a specificity of 100% and a sensitivity of 100%.
