A CHROMOSOMAL INTEGRATION SYSTEM FOR STABILIZATION OF HETEROLOGOUS GENES IN SALMONELLA BASED VACCINE STRAINS
A CHROMOSOMAL INTEGRATION SYSTEM FOR STABILIZATION OF HETEROLOGOUS GENES IN SALMONELLA BASED VACCINE STRAINS
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DOI：
10.1016/0882-4010(88)90002-2
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发表时间：
1988-12-01
影响因子：
3.8
通讯作者：
HACKETT, J
中科院分区：
文献类型：
--
作者：
HONE, D;ATTRIDGE, S;HACKETT, J
我们开发了一种系统，编码来自肠道病原体的抗原的异源DNA可以重组到减毒沙门氏菌携带菌株的染色体中。该系统涉及两个步骤：（i）将 hisOG 缺失突变整合到染色体中； (ii)用完整的hisOG区域和编码目的抗原的异源DNA片段替换hisOG缺失。可以选择重组体(his+)。该系统用于将产肠毒素大肠杆菌的 K88 菌毛编码基因整合到鼠伤寒沙门氏菌 galE 突变体 (LT2H1) 的染色体中。重组体以1.0次之间的频率被检测到。 10-3 和 1.5 倍。 10-3。多种测试证实K88基因整合到LT2H1的染色体上并表达。重组体的稳定性在体内和体外进行了测试。当给小鼠口服给药时，重组体引发了针对 K88 的血清抗体反应，并保留了载体菌株的沙门氏菌疫苗潜力。
We have developed a system whereby heterologous DNA encoding an antigen from an enteropathogen may be recombined into the chromosome of an attenuated Salmonella carrier strain. The system involves two steps: (i) integration of a hisOG deletion mutation into the chromosome; (ii) replacement of the hisOG deletion by the complete hisOG region and the segment of heterologous DNA which encodes the antigen of interest. Recombinants may be selected (his+). The system was used to integrate the genes encoding K88 fimbriae from enterotoxigenic Escherichia coli into the chromosome of a galE mutant of Salmonella typhimurium (LT2H1). Recombinants were detected at a frequency of between 1.0 .times. 10-3 and 1.5 .times. 10-3. A variety of tests confirmed that the K88 genes were integrated into the chromosome of LT2H1 and were expressed. The stability of the recombinant was tested both in vivo and in vitro. When administered orally to mice, the recombinant elicited a serum antibody response to K88, and retained the Salmonella vaccine potential of the vector strain.