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卡尔斯伯酵母MAL6位点麦芽糖酶基因的一级结构

Primary structure of the maltase gene of the MAL6 locus of Saccharomyces carlsbergensis.

作者信息

Hong S H, Marmur J

出版信息

Gene. 1986;41(1):75-84. doi: 10.1016/0378-1119(86)90269-6.

Abstract

We have determined the complete nucleotide (nt) sequence of a 2937-bp DNA fragment containing the yeast maltase (EC 3.2.1.20) gene (MAL6S) as well as part of the contiguous maltose permease gene (MAL6T) from the MAL6 locus of Saccharomyces carlsbergensis. The MAL6S gene encodes an alpha-glucosidase that is required for the utilization of maltose as a carbon source by yeast. The 5' transcription initiation sites for both MAL6S and MAL6T were determined by primer extension experiments using reverse transcriptase. The sequence data show one major open reading frame (ORF) of 584 amino acids (aa) for maltase with a calculated Mr of 68 107, somewhat larger than the value of 63 000 previously determined by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) analysis. The nucleotide sequences upstream of both the MAL6S and MAL6T genes, which are divergently transcribed, show common structural features for the transcription initiation of yeast genes as well as signals required for their translation. The codon bias index shows that the MAL6S gene is moderately expressed. The possible significance of two 17-bp dyad symmetric sequences, found in the intergenic region of MAL6S and MAL6T, for the control of expression of these genes is also discussed.

摘要

我们已经确定了一个2937bp DNA片段的完整核苷酸(nt)序列,该片段包含来自卡尔斯伯酵母MAL6位点的酵母麦芽糖酶(EC 3.2.1.20)基因(MAL6S)以及相邻的麦芽糖通透酶基因(MAL6T)的一部分。MAL6S基因编码一种α-葡萄糖苷酶,酵母利用麦芽糖作为碳源时需要该酶。通过使用逆转录酶的引物延伸实验确定了MAL6S和MAL6T的5'转录起始位点。序列数据显示麦芽糖酶有一个由584个氨基酸(aa)组成的主要开放阅读框(ORF),计算所得的分子量为68107,略大于先前通过十二烷基硫酸钠(SDS)-聚丙烯酰胺凝胶电泳(PAGE)分析确定的63000的值。MAL6S和MAL6T基因上游的核苷酸序列呈反向转录,显示出酵母基因转录起始的共同结构特征以及它们翻译所需的信号。密码子偏好指数表明MAL6S基因表达适度。还讨论了在MAL6S和MAL6T基因间区域发现的两个17bp的二元对称序列对这些基因表达调控的可能意义。

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