Graves M C, Mullenbach G T, Rabinowitz J C
Proc Natl Acad Sci U S A. 1985 Mar;82(6):1653-7. doi: 10.1073/pnas.82.6.1653.
We have constructed a library of Clostridium pasteurianum DNA cloned in the plasmid pBR322. Based on the known amino acid sequence for C. pasteurianum ferredoxin, a 64-fold degenerate heptadecanucleotide pool was synthesized. This mixed probe hybridized to two clones which were shown to contain greater than 6 kilobase pairs of the same genomic DNA. Sequence analysis of a common Sau3A1 0.6-kilobase-pair fragment revealed that it contains the information for the apoferredoxin structural gene. According to the DNA sequence, the only post-translational processing of this small apoprotein is the hydrolysis of the initiator methionine. Putative transcription and translation start and stop signals are present within the sequence.
我们构建了一个克隆于质粒pBR322的巴氏梭菌DNA文库。基于巴氏梭菌铁氧化还原蛋白已知的氨基酸序列,合成了一个64倍简并的十七聚核苷酸库。这个混合探针与两个克隆杂交,这两个克隆被证明含有大于6千碱基对的相同基因组DNA。对一个常见的Sau3A1 0.6千碱基对片段的序列分析表明,它包含脱辅基铁氧化还原蛋白结构基因的信息。根据DNA序列,这种小脱辅基蛋白唯一的翻译后加工是起始甲硫氨酸的水解。推测的转录和翻译起始及终止信号存在于该序列中。