Nudel U, Soreq H, Littauer U Z
Eur J Biochem. 1976 Apr 15;64(1):115-21. doi: 10.1111/j.1432-1033.1976.tb10279.x.
Rabbit globin mRNA species containing poly(A) segments of different lengths were prepared by partial phosphorolysis of mRNA with Escherichia coli polynucleotide phosphorylase. By varying the salt concentration and the time of incubation of the phosphorolysis mixture, as well as performing oligo(dT)-cellulose chromatography at 22 degrees C and at 4 degrees C, globin mRNA preparations containing poly(A) segments of approximately 122, 95, 68, 39, 32, 21, and 16 adenylate residues were obtained. It was found that the functional stability of the mRNA species containing 32 or more adenylate residues after injection into Xenopus laevis oocytes equaled that of the native globin mRNA. On the other hand, the functional stability of mRNA containing an average number of 21 adenylate residues was about 30% of the native mRNA, while that of mRNA containing 16 adenylate residues was as low as poly(A)-free globin MRNA.
通过用大肠杆菌多核苷酸磷酸化酶对mRNA进行部分磷酸解反应,制备了含有不同长度聚(A)片段的兔珠蛋白mRNA种类。通过改变磷酸解混合物的盐浓度和温育时间,以及在22℃和4℃下进行寡聚(dT)-纤维素层析,获得了含有大约122、95、68、39、32、21和16个腺苷酸残基的聚(A)片段的珠蛋白mRNA制剂。结果发现,注射到非洲爪蟾卵母细胞后,含有32个或更多腺苷酸残基的mRNA种类的功能稳定性与天然珠蛋白mRNA的功能稳定性相当。另一方面,平均含有21个腺苷酸残基的mRNA的功能稳定性约为天然mRNA的30%,而含有16个腺苷酸残基的mRNA的功能稳定性则与无聚(A)的珠蛋白mRNA一样低。