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热带假丝酵母过氧化物酶体蛋白编码基因的高水平表达及分子克隆

High-level expression and molecular cloning of genes encoding Candida tropicalis peroxisomal proteins.

作者信息

Kamiryo T, Okazaki K

出版信息

Mol Cell Biol. 1984 Oct;4(10):2136-41. doi: 10.1128/mcb.4.10.2136-2141.1984.

DOI:10.1128/mcb.4.10.2136-2141.1984
PMID:6504042
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC369032/
Abstract

The development of peroxisomes in the cells of Candida tropicalis grown on oleic acid was accompanied by a markedly high expression of peroxisomal proteins. On the basis of this finding, the nuclear DNA library of this yeast was screened by differential hybridization, and 102 clones of oleic acid-inducible sequences were isolated. Seven coding regions were found to form clusters in three stretches of the genomic DNA. Five of the regions were identified as genes for peroxisomal polypeptides (PXPs). The coding sequence for PXP-2 hybrid selected an additional mRNA for PXP-4, the subunit of long-chain acyl coenzyme A oxidase, which was the most abundant PXP. PXP-2 and PXP-4 were close in apparent molecular weight and generated similar peptides when digested with a protease. The gene for PXP-4 was adjacent to that for PXP-2 on the genome and also hybridized to the mRNA coding for PXP-5. These and other similar results suggest that the genes for the peroxisomal proteins of this organism arose by duplication of a few ancestral genes.

摘要

在以油酸为碳源生长的热带假丝酵母细胞中，过氧化物酶体的发育伴随着过氧化物酶体蛋白的显著高表达。基于这一发现，通过差异杂交筛选了该酵母的核DNA文库，分离出102个油酸诱导序列的克隆。发现7个编码区在基因组DNA的三个片段中形成簇。其中5个区域被鉴定为过氧化物酶体多肽（PXP）的基因。PXP-2的编码序列选择了另一种PXP-4的mRNA，PXP-4是长链酰基辅酶A氧化酶的亚基，也是最丰富的PXP。PXP-2和PXP-4的表观分子量相近，用蛋白酶消化时产生相似的肽段。PXP-4基因在基因组上与PXP-2基因相邻，并且也与编码PXP-5的mRNA杂交。这些以及其他类似结果表明，该生物体过氧化物酶体蛋白的基因是由少数祖先基因的复制产生的。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/518e/369032/016d3cf6e530/molcellb00152-0207-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/518e/369032/f84f3a62ffb1/molcellb00152-0205-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/518e/369032/92719159adba/molcellb00152-0206-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/518e/369032/b1429c056291/molcellb00152-0207-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/518e/369032/016d3cf6e530/molcellb00152-0207-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/518e/369032/f84f3a62ffb1/molcellb00152-0205-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/518e/369032/92719159adba/molcellb00152-0206-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/518e/369032/b1429c056291/molcellb00152-0207-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/518e/369032/016d3cf6e530/molcellb00152-0207-b.jpg

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本文引用的文献

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Biosynthesis of peroxisomal enzymes in the methylotrophic yeast Hansenula polymorpha.甲醇营养型酵母异常汉逊酵母中过氧化物酶体酶的生物合成。
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Proc Natl Acad Sci U S A. 1986 Mar;83(5):1232-6. doi: 10.1073/pnas.83.5.1232.
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Complete nucleotide sequence of the peroxisomal acyl CoA oxidase from the alkane-utilizing yeast Candida maltosa.利用烷烃的酵母麦芽糖假丝酵母中过氧化物酶体酰基辅酶A氧化酶的完整核苷酸序列。
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Cloning of cDNA coding for peroxisomal acyl-CoA oxidase from the yeast Candida tropicalis pK233.从热带假丝酵母pK233中克隆编码过氧化物酶体酰基辅酶A氧化酶的cDNA
Proc Natl Acad Sci U S A. 1985 Jun;82(12):3973-7. doi: 10.1073/pnas.82.12.3973.
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KGB: a single buffer for all restriction endonucleases.KGB：所有限制性内切酶的单一缓冲液。
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Interspecific complementation analysis by protoplast fusion of Candida tropicalis and Candida albicans adenine auxotrophs.热带假丝酵母和白色念珠菌腺嘌呤营养缺陷型原生质体融合的种间互补分析。
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