Improved techniques for the isolation of intact thyroglobulin-synthesizing polysomes

✍ Scribed by Eric Davies; J.E. Dumont; G. Vassart

• Publisher: Elsevier Science
• Year: 1977
• Tongue: English
• Weight: 490 KB
• Volume: 80
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(77)90647-9

✦ Synopsis

A method is presented, based on the use of an extraction buffer containing 200 mM Tris-HCl, pH 8.5, which permits the isolation in high yield of apparently undegraded polysomes from beef thyroid. Upon treatment with detergent, both the 8OOg pellet and the 27,000g pellet yield polysome profiles exhibiting very few monosomes and small polysomes but a major, homogenous peak of very large, thyroglobulin-synthesizing polysomes. No such large polysomes were evident in gradients of free polysomes. The polysomal nature of this peak of very large polysomes was verified by treatment with detergent and ribonuclease, by electron microscopy, and by extraction of the mRNA.