Stachelek C, Stachelek J, Swan J, Botstein D, Konigsberg W
Nucleic Acids Res. 1986 Jan 24;14(2):945-63. doi: 10.1093/nar/14.2.945.
The hexokinase A (HKA) and hexokinase B (HKB) genes of Saccharomyces cerevisiae have been cloned from a library of yeast genomic DNA. Using an in vitro glucose phosphorylation assay, the HKB gene was located on a plasmid carrying a 13.6 kb fragment of yeast DNA. After subcloning the relevant restriction fragments, the nucleotide sequence of the HKB gene was determined. Using this information, we were able to locate the HKA gene on a plasmid carrying this gene, which we then sequenced. Approximately 43% of the amino acid sequence of HKB was determined directly from 24 tryptic peptides. The results are in complete agreement with those derived from the DNA sequence and are consistent with the results of x-ray crystallography. Comparison of the amino acid sequences of HKA and HKB show that 378 out of 485 residues are identical. The 5' flanking region of the A gene contains nucleotide sequences expected for genes that are expressed at relatively high levels in yeast. The 24 base pair hyphenated palindrome at the 3' end of the HKB gene may be a site for termination of transcription of this gene.
酿酒酵母的己糖激酶A（HKA）和己糖激酶B（HKB）基因已从酵母基因组DNA文库中克隆出来。通过体外葡萄糖磷酸化测定，HKB基因位于携带13.6 kb酵母DNA片段的质粒上。在亚克隆相关的限制性片段后，确定了HKB基因的核苷酸序列。利用这些信息，我们能够在携带该基因的质粒上定位HKA基因，然后对其进行测序。HKB约43%的氨基酸序列直接由24个胰蛋白酶肽段确定。结果与从DNA序列推导的结果完全一致，并且与X射线晶体学的结果相符。HKA和HKB氨基酸序列的比较表明，485个残基中有378个是相同的。A基因的5'侧翼区域包含酵母中相对高水平表达的基因所预期的核苷酸序列。HKB基因3'端的24个碱基对的连字符回文序列可能是该基因转录终止的位点。
