麦芽糖假丝酵母中POX2基因的克隆与特性分析

Cloning and characterization of the POX2 gene in Candida maltosa.

作者信息

Masuda Y, Park S M, Ohta A, Takagi M

机构信息

Department of Agricultural Chemistry, University of Tokyo, Japan.

出版信息

Gene. 1995 Dec 29;167(1-2):157-61. doi: 10.1016/0378-1119(95)00655-9.

DOI:10.1016/0378-1119(95)00655-9

PMID:8566769

Abstract

To study the function of acyl-CoA oxidase in an n-alkane-assimilating yeast, Candida maltosa, we isolated the POX2 gene which is a member of the acyl-CoA oxidase gene family. POX2 had a 2172-bp open reading frame (ORF) encoding an approx. 84-kDa polypeptide (724 amino acids (aa)) and was contiguous to POX4, another member of the acyl-CoA oxidase gene family on the same chromosomal DNA in a convergent arrangement. Northern blot analysis revealed that the expression of POX2 was induced in cells grown on oleic acid, n-tetradecanol and n-tetradecane. By using a gene-disruption technique, we constructed strains (termed P2DD and P4DD) in which both alleles of POX2 and POX4 were disrupted. The P2DD strain was normal in assimilation of various hydrophobic carbon sources, such as n-tetradecane, n-tetradecanol and oleic acid. In contrast, the P4DD strain was defective in its ability to grow on such hydrophobic carbon sources.

摘要

为了研究酰基辅酶A氧化酶在同化正构烷烃的酵母——麦芽糖假丝酵母中的功能，我们分离了POX2基因，它是酰基辅酶A氧化酶基因家族的成员之一。POX2有一个2172 bp的开放阅读框（ORF），编码一个约84 kDa的多肽（724个氨基酸（aa）），并且在同一染色体DNA上以反向排列与酰基辅酶A氧化酶基因家族的另一个成员POX4相邻。Northern印迹分析表明，POX2的表达在以油酸、正十四烷醇和正十四烷为碳源生长的细胞中被诱导。通过基因破坏技术，我们构建了POX2和POX4两个等位基因均被破坏的菌株（称为P2DD和P4DD）。P2DD菌株在同化各种疏水碳源（如正十四烷、正十四烷醇和油酸）方面正常。相比之下，P4DD菌株在利用此类疏水碳源生长的能力上存在缺陷。