狂犬病毒核蛋白在昆虫细胞中的表达及免疫原性的研究

目的在昆虫细胞中表达出狂犬病毒核蛋白(NP),并探讨重组NP的免疫原性.方法采用杆状病毒表达系统在Sf9昆虫细胞中表达狂犬病毒核蛋白,用直接免疫荧光(DFA)、SDS-PAGE和Western blot对表达产物进行检测和分析,以感染重组杆状病毒的昆虫细胞裂解液免疫小鼠,检测表达产物的免疫原性.结果重组杆状病毒在昆虫细胞中获得表达,免疫小鼠可产生抗核蛋白抗体.结论在Sf9昆虫细胞中表达出狂犬病毒核蛋白,重组产物具有生物活性.     

Biochemical aspects of radiation biology

Summary In order to analyze the mechanisms of biological radiation effects, the events after radiation energy absorption in irradiated organisms have to be studied by physico-chemical and biochemical methods. The radiation effects in vitro on biomolecules, especially DNA, are described, as well as their alterations in irradiated cells. Whereas in vitro, in aqueous solution, predominantly OH radicals are effective and lead to damage in single moieties of the DNA, in vivo the direct absorption of radiation energy leads to locally multiply-damaged sites, which produce DNA double-strand breaks and locally denatured regions. DNA damage will be repaired in irradiated cells. Error free repair leads to the original nucleotide sequence in the genome by excision or by recombination. Error prone repair (mutagenic repair), leads to mutation. However, the biochemistry of these processes, regulated by a number of genes, is poorly understood. In addition, more complex reactions, such as gene amplification and transposition of mobile gene elements, are responsible for mutation or malignant transformation.     

汉滩病毒S基因0.7kb片段与M基因G2片段不同拼接方式嵌合基因原核表达产物的初步比较

比较汉滩病毒S、M基因部分片段的嵌合基因不同拼接方式表达产物的活性。构建了嵌合基因原核表达载体pGEX-4T-1-S0.7G2,并与已构建的pGEX-4T-1-G2S0.7的诱导表达产物进行了比较。结果表明,融合蛋白同时保持亲本蛋白的结合活性,且前者表达产物的结合活性始终比后者低。研究表明,嵌合基因的不同拼接方式对融合蛋白的活性可能有一定的影响。