Hanafusa T, Saito K, Tominaga A, Enomoto M
Department of Biology, Faculty of Science, Okayama University, Japan.
Mol Gen Genet. 1993 Jan;236(2-3):260-6. doi: 10.1007/BF00277121.
The nucleotide sequence of Salmonella abortus-equi fljA, which together with the phase 2 flagellin gene constitutes the fljBA operon and encodes the repressor for the phase 1 flagellin gene fliC, was determined. The repressor was predicted to be a basic protein consisting of 179 amino acid residues (M(r) = 20419 Da) encoded by ORFII. This was confirmed by the fact that host fliC is repressed by plasmid-encoded ORFII, which indeed expresses a 20 kDa product as determined by urea SDS-polyacrylamide gel electrophoresis. An amino acid sequence capable of forming a helix-turn-helix type of structure was predicted in the C-terminal region of FljA. A rho-independent intercistronic terminator was detected between fljB and fljA. Chloramphenicol acetyltransferase (CAT) assays of fusions indicated that the terminator is capable of reducing expression of fljA to the level of a few percent, relative to fljB in broth cultures and to 1% in M9 glycerol cultures.
测定了马流产沙门氏菌fljA的核苷酸序列,它与2期鞭毛蛋白基因共同构成fljBA操纵子,并编码1期鞭毛蛋白基因fliC的阻遏物。预测该阻遏物是一种由179个氨基酸残基组成的碱性蛋白(M(r)=20419 Da),由ORFII编码。这一点得到了证实,即宿主fliC被质粒编码的ORFII抑制,通过尿素SDS-聚丙烯酰胺凝胶电泳测定,ORFII确实表达了一种20 kDa的产物。在FljA的C端区域预测到一个能够形成螺旋-转角-螺旋结构类型的氨基酸序列。在fljB和fljA之间检测到一个不依赖ρ因子的顺反子间终止子。融合体的氯霉素乙酰转移酶(CAT)分析表明,相对于肉汤培养物中的fljB和M9甘油培养物中的1%,该终止子能够将fljA的表达水平降低到百分之几。