通过定量免疫沉淀法测定非洲爪蟾卵母细胞中晶状体晶状体蛋白的合成。

Synthesis of lens crystallins in Xenopus oocytes as determined by quantitative immunoprecipitation.

作者信息

Asselbergs F A, Van Venrooij W J, Bloemendal H

出版信息

Eur J Biochem. 1978 Jul 3;87(3):517-24. doi: 10.1111/j.1432-1033.1978.tb12402.x.

DOI:10.1111/j.1432-1033.1978.tb12402.x

Abstract

Total poly(A)-containing calf lens mRNA was microinjected into Xenopus oocytes and synthesis of alpha, beta, and gamma-crystallins was demonstrated. By a method of quantitative immunoprecipitation the rate of translation of purified 14S alphaA2-crystallin mRNA was compared with translation of 9-S rabbit globin mRNA. Maximal response of oocytes was obtained with virtually the same molar amounts of mRNA, taking into account the larger size of the alphaA2-crystallin mRNA. Kinetics of translation were also very similar and both mRNAs were translated with similar rate and efficiency for at least two days. It was estimated that 20-30 polypeptide chains per hour per mRNA molecule were synthesized.

摘要

将含poly(A)的小牛晶状体总mRNA显微注射到非洲爪蟾卵母细胞中，并证实了α、β和γ晶状体蛋白的合成。通过定量免疫沉淀法，将纯化的14SαA2-晶状体蛋白mRNA的翻译速率与9S兔珠蛋白mRNA的翻译速率进行了比较。考虑到αA2-晶状体蛋白mRNA的较大尺寸，用几乎相同摩尔量的mRNA可获得卵母细胞的最大反应。翻译动力学也非常相似，两种mRNA至少在两天内以相似的速率和效率进行翻译。据估计，每个mRNA分子每小时合成20 - 30条多肽链。