Johnston T C, Thompson R B, Baldwin T O
J Biol Chem. 1986 Apr 15;261(11):4805-11.
The nucleotide sequence of the 1.30-kilobase EcoRI/BglII fragment from Vibrio harveyi carrying the majority of the luciferase beta subunit coding region (luxB gene) has been determined. The EcoRI/BglII fragment was derived from a 4.0-kilobase HindIII fragment carrying both luxA and luxB which was detected in a genomic clone bank based on the expression of bioluminescence from colonies of Escherichia coli carrying V. harveyi HindIII fragments in plasmid pBR322 (Baldwin, T. O., Berends, T., Bunch, T. A., Holzman, T. F., Rausch, S. K., Shamansky, L., Treat, M. L., and Ziegler, M. M. (1984) Biochemistry 23, 3663-3667). The entire alpha subunit coding sequence (luxA gene) and the amino-terminal 13 codons of the beta subunit sequence (luxB gene) were contained on a 1.85-kilobase EcoRI fragment, the sequence of which has been reported (Cohn, D. H., Mileham, A. J., Simon, M. I., Nealson, K. H., Rausch, S. K., Bonam, D., and Baldwin, T. O. (1985) J. Biol. Chem. 260, 6139-6146). The beta subunit coding sequence was found to terminate 972 bases past the start of the luxB coding sequence. The beta subunit had a calculated molecular weight of 36,349 and comprised a total of 324 amino acid residues; the alpha beta dimer had a molecular weight (alpha + beta) of 76,457. There were 27 base pairs separating the stop codon of the beta subunit structural gene and a 340-base open reading frame extending to (and beyond) the distal BglII site. Approximately two-thirds of the beta subunit was sequenced by protein chemical techniques. The amino acid sequence predicted from the DNA sequence, with few exceptions, confirmed the chemically determined sequence, and the measured amino acid composition was in excellent agreement with the composition implied from the DNA sequence.
已确定来自哈维氏弧菌的携带大部分荧光素酶β亚基编码区（luxB基因）的1.30千碱基EcoRI/BglII片段的核苷酸序列。该EcoRI/BglII片段源自一个4.0千碱基的HindIII片段，该片段同时携带luxA和luxB，这是在一个基因组克隆文库中检测到的，该文库基于携带哈维氏弧菌HindIII片段的大肠杆菌菌落（位于质粒pBR322中）的生物发光表达（鲍德温，T.O.，贝伦兹，T.，邦奇，T.A.，霍尔兹曼，T.F.，劳施，S.K.，沙曼斯基，L.，特里特，M.L.，和齐格勒，M.M.（1984年）《生物化学》23卷，3663 - 3667页）。整个α亚基编码序列（luxA基因）和β亚基序列的氨基末端13个密码子（luxB基因）包含在一个1.85千碱基的EcoRI片段上，其序列已被报道（科恩，D.H.，米勒姆，A.J.，西蒙，M.I.，尼尔森，K.H.，劳施，S.K.，博纳姆，D.，和鲍德温，T.O.（1985年）《生物化学杂志》260卷，6139 - 6146页）。发现β亚基编码序列在luxB编码序列起始点之后972个碱基处终止。β亚基的计算分子量为36,349，总共由324个氨基酸残基组成；αβ二聚体的分子量（α + β）为76,457。在β亚基结构基因的终止密码子和一个延伸至（并超出）远端BglII位点的340碱基开放阅读框之间有27个碱基对。大约三分之二的β亚基是通过蛋白质化学技术测序的。从DNA序列预测的氨基酸序列，除少数例外，证实了化学测定的序列，并且测量的氨基酸组成与从DNA序列推断的组成非常一致。
