文昌鱼卵巢含Poly(A)的RNA的分离纯化

文昌鱼是无脊椎动物和脊椎动物之间的过渡类型,是研究动物演化的理想材料.了解文昌鱼在代谢过程中发生的生物化学变化和从分子生物学观点研究个体发育的特点,在科学上是有重要价值的.这必将受到胚胎学、细胞学、遗传学和生理学工作者的极大重视. 文昌鱼的卵母细胞是一种生长非常旺盛的生殖细胞.

ISOLATION AND PURIFICATION OF OVARIAN POLYADENYL-ATED RNA FROM AMPHIOXUS

Amphioxus, a transitory form between the invertebrate and the vertebrate, is an ideal material for the study of animal evolution. The material used in this experiment is the common Amphioxus collected from Qingdao, Branchiosloma belcheri tsinytdoense Tchang et Koo.The purpose of this study is to find out the biochemical changes taking place in the metabolic processes of this animal and to demonstrate from the standpoint of molecular biology the speciality of its ontogenetic development. By means of ultracentrifugal technique and Oligo (dT)-cellulose affinity chromatography, ovarian Poly (A) RNA from Branchiostorna belcheri tsingtaoense Tehang et Koo was isolated and purified,The polysomes were obtained from the supernatant of the oocyte lysate through the ultracentrifugation of 1.0M sucrose solution and the total polysomal RNA was extracted from the polysome precipitate by phenol-chloroform-isoamyl alcohol (50:48:2). Poly (A) RNA. in the ovarian eggs of Amphioxus was purified by Oligo (dT)-cellulose chromatography to remove rRNA. Finally, Poly(A) RNA was precipitated by 2 volumes of ethanol and washed,with 4M LiCL and 3:1 ethanol saline to obtain Poly(A)RNA with biological activity. The recovery rate of the total polysomal RNA is 285μg per ml homogenate of the ovarian eggs.The mRNA-containing Poly(A),purified by means of Oligo(dT)-cellulose is 5.2% of the total polysomal RNA and 4% of the total RNA.The Polp (A)RNA obtained shows the typical absorption spectrum of RNA in ultraviolet spectral analysis.