Minton N P, Atkinson T, Bruton C J, Sherwood R F
Gene. 1984 Nov;31(1-3):31-8. doi: 10.1016/0378-1119(84)90192-6.
The complete nucleotide sequence of the Pseudomonas chromosomal gene coding for the enzyme carboxypeptidase G2 (CPG2) has been determined. The nucleotide sequence obtained has been confirmed by comparing the predicted amino acid sequence with that of randomly derived peptide fragments and by N-terminal sequencing of the purified protein. The gene has been shown to code for a 22 amino acid signal peptide at its N-terminus which closely resembles the signal peptides of other secreted proteins. An alternative 36 amino acid signal peptide which may function in Pseudomonas has also been identified. The codon utilisation of the gene is influenced by the high G + C (67.2%) content of the DNA and exhibits a 92.8% preference for codons ending in G or C. This unusual codon preference may contribute to the generally observed weak expression of Pseudomonas genes in Escherichia coli. A region of DNA upstream of the structural gene has also been sequenced and a ribosome binding site and two putative promoter sequences identified.
已确定编码羧肽酶G2(CPG2)的假单胞菌染色体基因的完整核苷酸序列。通过将预测的氨基酸序列与随机获得的肽片段的序列进行比较,并对纯化蛋白进行N端测序,证实了所获得的核苷酸序列。该基因在其N端编码一个22个氨基酸的信号肽,该信号肽与其他分泌蛋白的信号肽非常相似。还鉴定出了一个可能在假单胞菌中起作用的36个氨基酸的替代信号肽。该基因的密码子使用受DNA高G+C(67.2%)含量的影响,对以G或C结尾的密码子表现出92.8%的偏好。这种不寻常的密码子偏好可能导致假单胞菌基因在大肠杆菌中普遍观察到的弱表达。还对结构基因上游的一段DNA区域进行了测序,并鉴定出一个核糖体结合位点和两个假定的启动子序列。