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[编码软骨蛋白聚糖核心蛋白的cDNA克隆的构建]

[Construction of cDNA clones encoding for cartilage proteoglycan core protein].

作者信息

Sai S

出版信息

Nihon Seikeigeka Gakkai Zasshi. 1987 Jan;61(1):63-73.

PMID:3572140
Abstract

In order to investigate cartilage proteoglycan mRNAs and genes, a cDNA expression library was constructed from chick embryonic sternal mRNAs, and cDNA clones encoding for chondroitin sulfate proteoglycan core protein were screened from the library. Total RNA extracted from chick embryonic sterna was translated in a cell-free system using rabbit reticulocyte lysate. Immunoprecipitation of products of cell-free translation using antiserum against cartilage proteoglycan yielded a component of about 340 kd polypeptide. This polypeptide is most likely the translational product of proteoglycan core protein mRNA. A chicken embryo sternal cartilage cDNA library, created in the plasmid expression vector pUC 9, was screened for sequences coding for immunologically detectable core protein, and two cDNA clones were isolated. Northern blot hybridization of sternal cartilage RNA revealed a single mRNA of about 8.1 kb for proteoglycan core protein.

摘要

为了研究软骨蛋白聚糖mRNA和基因,从鸡胚胸骨mRNA构建了一个cDNA表达文库,并从该文库中筛选出编码硫酸软骨素蛋白聚糖核心蛋白的cDNA克隆。从鸡胚胸骨中提取的总RNA在无细胞系统中用兔网织红细胞裂解物进行翻译。用抗软骨蛋白聚糖抗血清对无细胞翻译产物进行免疫沉淀,得到一种约340kd多肽的成分。该多肽很可能是蛋白聚糖核心蛋白mRNA的翻译产物。在质粒表达载体pUC 9中构建的鸡胚胸骨软骨cDNA文库,筛选编码免疫可检测核心蛋白的序列,分离出两个cDNA克隆。胸骨软骨RNA的Northern印迹杂交显示蛋白聚糖核心蛋白有一条约8.1kb的单一mRNA。

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