Identification, cloning and sequence determination of the genes specifying hexokinase A and B from yeast.
Identification, cloning and sequence determination of the genes specifying hexokinase A and B from yeast.
复制标题
酵母中己糖激酶 A 和 B 基因的鉴定、克隆和序列测定。
DOI：
10.1093/nar/14.2.945
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发表时间：
1986
影响因子：
14.9
通讯作者：
Konigsberg,W
中科院分区：
文献类型：
--
作者：
Stachelek,C;Stachelek,J;Swan,J;Botstein,D;Konigsberg,W
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 DHA 文库中克隆出来。使用体外葡萄糖磷酸化测定，将 HKB 基因定位在携带 13.6 kb 酵母 DHA 片段的质粒上。亚克隆相关限制性片段后，确定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 大约 43% 的氨基酸序列是直接从 24 种胰蛋白酶肽中确定的。结果与 DHA 序列得出的结果完全一致，并且与 X 射线晶体学的结果一致。 HKA和HKB的氨基酸序列的比较显示485个残基中有378个是相同的。 A 基因的 5' 侧翼区域包含预期在酵母中以相对高水平表达的基因的核苷酸序列。 HKB基因3'端的24个碱基对连字符回文可能是该基因转录的终止位点。
The hexokinase A (HKA) and hexokinase B (HKB) genes of Saccharomyces cerevisiae have been cloned from a library of yeast genomic DHA. Using aninvitroglucose phosphorylation assay, the HKB gene was located on a plasmid carrying a 13.6 kb fragment of yeast DHA. 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 seqoenced. 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 DHA 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.