核糖核酸酶U2：克隆、在毕赤酵母中表达及活性重组蛋白的亲和层析纯化

Ribonuclease U2: cloning, production in Pichia pastoris and affinity chromatography purification of the active recombinant protein.

作者信息

Martínez-Ruiz A, García-Ortega L, Kao R, Oñaderra M, Mancheño J M, Davies J, Martínez del Pozo A, Gavilanes J G

机构信息

Departmento de Bioquimica y Biologia Molecular I, Facultad de Quimica, Universidad Complutense, Madrid, Spain.

出版信息

FEMS Microbiol Lett. 2000 Aug 15;189(2):165-9. doi: 10.1111/j.1574-6968.2000.tb09224.x.

DOI:10.1111/j.1574-6968.2000.tb09224.x

PMID:10930732

Abstract

RNase U2 is an endoribonuclease secreted by the fungus Ustilago sphaerogena. Its genomic DNA (rnu2), containing an intron of 116 bp, has been isolated and cloned. The corresponding cDNA has also been synthesized. The recombinant RNase U2 was successfully produced in Pichia pastoris, fused to the yeast alkaline phosphatase signal peptide. The recombinant RNase U2, purified by affinity chromatography, contains three extra amino acids at its amino-terminal end and retains the enzymatic and spectroscopic properties of the natural fungal protein.

摘要

核糖核酸酶U2是由球形黑粉菌分泌的一种核糖核酸内切酶。其基因组DNA（rnu2）含有一个116 bp的内含子，已被分离和克隆。相应的cDNA也已合成。重组核糖核酸酶U2在毕赤酵母中成功产生，与酵母碱性磷酸酶信号肽融合。通过亲和层析纯化的重组核糖核酸酶U2在其氨基末端含有三个额外的氨基酸，并保留了天然真菌蛋白的酶学和光谱学性质。