Norton J M, Low J M, Klotz M G
Department of Plants, Soils and Biometerology, Utah State University, Logan 84322-4820, USA.
FEMS Microbiol Lett. 1996 Jun 1;139(2-3):181-8. doi: 10.1111/j.1574-6968.1996.tb08200.x.
The gene encoding ammonia monooxygenase subunit A (AmoA) was found in three copies of the genome of the chemolithotrophic soil bacterium, Nitrosospira sp. NpAV. The open reading frame and flanking regions of the three copies were isolated from digested size fractionated genomic DNA using oligodeoxyribonucleotide primers and polymerase chain reaction. The three gene copies of amoA were sequenced and the sequences compared to each other. The open reading frames and the upstream and downstream flanking regions were nearly identical in the three copies. All three copies were expressed in recombinant Escherichia coli strains from the indigenous promoter producing a product of approximately 30 kDa. All amoA copies encode 274 amino acid polypeptides which have similarity to the ammonia monooxygenase acetylene-binding protein from Nitrosomonas europaea.
在化能自养土壤细菌亚硝化螺菌属NpAV的基因组中发现了编码氨单加氧酶亚基A(AmoA)的基因,共有三个拷贝。使用寡脱氧核糖核苷酸引物和聚合酶链反应从消化后的大小分级基因组DNA中分离出这三个拷贝的开放阅读框及其侧翼区域。对amoA的三个基因拷贝进行测序,并将序列相互比较。这三个拷贝中的开放阅读框以及上游和下游侧翼区域几乎相同。所有三个拷贝都在重组大肠杆菌菌株中从天然启动子表达,产生约30 kDa的产物。所有amoA拷贝都编码274个氨基酸的多肽,这些多肽与欧洲亚硝化单胞菌的氨单加氧酶乙炔结合蛋白相似。