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丝心蛋白信使核糖核酸的大小及其聚腺苷酸含量。

The size of fibroin messenger RNA and its polyadenylic acid content.

作者信息

Lizardi P M, Williamson R, Brown D D

出版信息

Cell. 1975 Mar;4(3):199-205. doi: 10.1016/0092-8674(75)90168-3.

Abstract

The molecular weight of fibroin messenger RNA has been measured by its contour length in the electron microscope and its electrophoretic mobility under conditions of complete denaturation. These methods give values of 5.6 times 10-6 and 6.0 times 10-6 daltons, respectively. The average molecular weight of 5.8 times 10-6 (1.6 times 10-4 base residues) would be expected to code for a protein of 414,000 daltons, a value about 12 percent larger than the most recently published size for fibroin. When the mRNA is prepared carefully to avoid shear breakage, it is as homogeneous in size as ribosomal RNA. Between 43 percent and 81 percent of the mRNA molecules bind to oligo(dT)-cellulose and are presumed to contain a poly(A) sequence. This sequence, isolated after ribonuclease digestion, comprises about 0.6 percent of the molecule, or an estimated 100 nucleotides. The poly(A) has been localized at the end of the molecule by hybridizing with a small poly (dT:dA) piece and subsequent visualization by electron microscopy. This observation, along with the ability of reverse transcriptase to transcribe fibroin mRNA with an oligo(dT) primer, demonstrates that the poly(A) is at the 3' end of the mRNA.

摘要

通过电子显微镜下的轮廓长度及其在完全变性条件下的电泳迁移率,已测定了丝心蛋白信使核糖核酸(mRNA)的分子量。这些方法分别给出了5.6×10⁻⁶和6.0×10⁻⁶道尔顿的值。预期平均分子量5.8×10⁻⁶(1.6×10⁻⁴个碱基残基)编码一种414,000道尔顿的蛋白质,该值比最近公布的丝心蛋白大小约大12%。当小心制备mRNA以避免剪切断裂时,其大小与核糖体RNA一样均一。43%至81%的mRNA分子与寡聚(dT)-纤维素结合,推测含有聚(A)序列。经核糖核酸酶消化后分离出的该序列约占分子的0.6%,或估计为100个核苷酸。通过与一小段聚(dT:dA)杂交并随后通过电子显微镜观察,已将聚(A)定位在分子的末端。这一观察结果,连同逆转录酶用寡聚(dT)引物转录丝心蛋白mRNA的能力,证明聚(A)位于mRNA的3'端。

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