Kliachko E V, Lysenko E S, Eremashvili M R, Kozlov Iu I, Strongin A Ia
Mol Biol (Mosk). 1988 May-Jun;22(3):760-6.
The half lives of mRNA for Escherichia coli chloramphenicol-acetyltransferase, Bacillus amyloliquefaciens alpha-amylase and human leucocyte interferon were measured in E. coli cells by molecular RNA.DNA hybridization. The effect of mutation in pnp gene, coding polynucleotide phosphorylase, on the stability of these mRNA was studied. The half life of interferon mRNA increases from 25 to 90 s in the pnp mutant, resulting in an increase of interferon accumulation. The stability of interferon in E. coli cells depends on the htpR gene, controlling the heat shock response. The yields of leucocyte interferons alpha-2, alpha I-1 and fibroblast interferon beta increase ten times in htpR mutants. Thus, by using pnp and htpR mutants it is possible to enhance considerably the eukaryotic gene expression in bacterial cells.
通过分子RNA-DNA杂交技术测定了大肠杆菌氯霉素乙酰转移酶、解淀粉芽孢杆菌α淀粉酶和人白细胞干扰素的信使核糖核酸（mRNA）半衰期。研究了编码多核苷酸磷酸化酶的pnp基因突变对这些mRNA稳定性的影响。在pnp突变体中，干扰素mRNA的半衰期从25秒增加到90秒，导致干扰素积累增加。大肠杆菌细胞中干扰素的稳定性取决于控制热休克反应的htpR基因。在htpR突变体中，白细胞干扰素α-2、αI-1和成纤维细胞干扰素β的产量增加了10倍。因此，通过使用pnp和htpR突变体，可以显著提高真核基因在细菌细胞中的表达。
