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酿酒酵母的组氨酸通透酶基因(HIP1)。

The histidine permease gene (HIP1) of Saccharomyces cerevisiae.

作者信息

Tanaka J, Fink G R

出版信息

Gene. 1985;38(1-3):205-14. doi: 10.1016/0378-1119(85)90219-7.

Abstract

The histidine-specific permease gene (HIP1) of Saccharomyces cerevisiae has been mapped, cloned, and sequenced. The HIP1 gene maps to the right arm of chromosome VII, approx. 11 cM distal to the ADE3 gene. The gene was isolated as an 8.6-kb BamHI-Sau3A fragment by complementation of the histidine-specific permease deficiency in recipient yeast cells. We sequenced a 2.4-kb subfragment of this BamHI-Sau3A fragment containing the HIP1 gene and identified a 1596-bp open reading frame (ORF). We confirmed the assignment of the 1596-bp ORF as the HIP1 coding sequence by sequencing a hip1 nonsense mutation. Analysis of the amino acid (aa) sequence of the HIP1 gene reveals several hydrophobic stretches, but shows no obvious N-terminal signal peptide. We have constructed a deletion of the HIP1 gene in vitro and replaced the wild-type copy of the gene with this deletion. The hip1 deletion mutant can grow when it is supplemented with 30 mM histidine, 50 times the amount required for the growth of HIP1 cells. Revertants of this deletion mutant able to grow on a normal level of histidine arise by mutation in unlinked genes. Both these observations suggest that there are additional, low-affinity pathways for histidine uptake.

摘要

酿酒酵母的组氨酸特异性通透酶基因(HIP1)已被定位、克隆和测序。HIP1基因定位于染色体VII的右臂,距离ADE3基因约11厘摩远。通过互补受体酵母细胞中组氨酸特异性通透酶缺陷,该基因作为一个8.6 kb的BamHI - Sau3A片段被分离出来。我们对这个包含HIP1基因的BamHI - Sau3A片段的一个2.4 kb亚片段进行了测序,并鉴定出一个1596 bp的开放阅读框(ORF)。通过对一个hip1无义突变进行测序,我们证实了这个1596 bp的ORF就是HIP1编码序列。对HIP1基因的氨基酸(aa)序列分析揭示了几个疏水区域,但未显示出明显的N端信号肽。我们在体外构建了HIP1基因的缺失,并将该基因的野生型拷贝替换为这个缺失。当补充30 mM组氨酸时,hip1缺失突变体能够生长,这是HIP1细胞生长所需量的50倍。这个缺失突变体的回复突变体能够在正常水平的组氨酸上生长,是通过非连锁基因的突变产生的。这两个观察结果都表明存在额外的、低亲和力的组氨酸摄取途径。

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