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目的 了解乙型肝炎病毒表面S +前S1融合重组DNA(乙型肝炎重组DNA)整合到中国地鼠卵巢细胞二氢叶酸还原酶阴性突变株细胞 (CHOdhfr- )染色体的变化及致瘤性。方法 应用分子克隆技术构建含HBsAg的表面抗原S +前S1基因的质粒经转染、克隆、加压 (MTX和MSX)筛选出高效表达乙型肝炎表面S +S1融合抗原的转基因细胞株 (GdSS1 18)进行染色体制片 ,同时将制备的细胞、细胞DNA和细胞匀浆皮下接种裸鼠观察 3周。结果 整合了外源乙型肝炎重组DNA的转基因细胞系 (GdSS1 18)不同的传代均发生染色体结构畸变 ,畸变率为 11% ,5 6%及 2 9% ,而未整合外源乙型肝炎重组DNA的对照CHOdhfr- 细胞为 6% ,二者的染色体众数无明显变化 ,均为 2 0条。整合与未整合外源重组DNA的细胞接种裸鼠以后观察 3周均无肿瘤生长。结论 整合有外源乙型肝炎重组DNA的细胞株 (GdSS1 18)其染色体结构畸变率高于未整合的CHO dhfr- 对照细胞 ,但二者的染色体众数仍为 2 0条 ;不同代数GdSS1 18细胞产物接种裸鼠后未见肿瘤的生长。
Objective To investigate the chromosomal changes and tumorigenicity of Sd pre-S1 fusion DNA (hepatitis B recombinant DNA) from Chinese hamster ovary (DHD) -reverse mutant CHO cells. Methods Plasmid containing S + pre-S1 gene of HBsAg was constructed by molecular cloning technology. Transgenic, GdSS1 (GdSS1) cells were screened by MTX and MSX. 18) Chromosome preparation, while the prepared cells, cell DNA and cell homogenate nude mice were inoculated subcutaneously for 3 weeks. Results Chromosome structural aberrations occurred in different passages of transgenic GdSS1 18 cells, which were integrated with exogenous hepatitis B recombinant DNA. The aberration rates were 11%, 56% and 29%, respectively. However, no integration of exogenous hepatitis B reorganization DNA control CHOdhfr- cells were 6%, both of the chromosome number no significant change, were 20. No tumor growth was observed after 3 weeks inoculation of nude mice with or without integration of exogenous recombinant DNA. Conclusion The chromosome aberration rate of GdSS1 18 cells with integrated recombinant DNA of hepatitis B is higher than that of unintegrated CHO dhfr-control cells, but the chromosome number of GdSS1 18 is still 20; GdSS1 18 Tumor growth was not seen after nude mice were inoculated with cell products.