Amandaraj M P, Roe B A
Biochemistry. 1975 Nov 18;14(23):5068-73. doi: 10.1021/bi00694a006.
By using column chromatography on varied media, the purification of several individual tRNAs from human placenta has been achieved. The crude human placenta tRNA was isolated using phenol extraction at pH 4.5 followed by DEAE-cellulose chromatography (B. Roe (1975), Nucleic Acids Res. 2, 21-42) and initially fractionated on BD-cellulose at neutral pH. Subsequent chromatography of the partially purified tRNA using high-speed, high-pressure liquid chromatography on RPC-5 and Aminex A-28 coupled with chromatography on BD-cellulose at acidic pH and on DEAE-Sephadex A-50 significantly shortened isolation time for milligram quantities of several pure tRNA species. Those tRNAs from human placenta obtained in a purity greater than 1.2 nmol/A260 unit are tRNAPhe, tRNAMet(i), tRNAVal(1a), tRNAVal(1b), and tRNAGly(1), while those obtained at purity of at least 0.8 nmol/A260 unit are tRNASer2 and tRNASer3. In addition, the use of Aminex A-28 as a chromatographic system for the isolation of tRNA is discussed.
通过在不同介质上进行柱色谱法，已实现从人胎盘中纯化出几种单个的转运RNA（tRNA）。人胎盘粗tRNA通过在pH 4.5下用苯酚萃取，随后进行DEAE - 纤维素色谱法（B. Roe（1975年），《核酸研究》2，21 - 42）进行分离，并首先在中性pH下于BD - 纤维素上分级分离。随后，使用RPC - 5和Aminex A - 28进行高速、高压液相色谱法对部分纯化的tRNA进行色谱分析，并结合在酸性pH下于BD - 纤维素上以及在DEAE - 葡聚糖A - 50上进行色谱分析，显著缩短了数毫克量的几种纯tRNA种类的分离时间。从人胎盘中获得的纯度大于1.2 nmol/A260单位的那些tRNA是苯丙氨酸tRNA（tRNAPhe）、起始甲硫氨酸tRNA（tRNAMet(i)）、缬氨酸tRNA(1a)（tRNAVal(1a)）、缬氨酸tRNA(1b)（tRNAVal(1b)）和甘氨酸tRNA(1)（tRNAGly(1)），而纯度至少为0.8 nmol/A260单位的那些是丝氨酸tRNA2（tRNASer2）和丝氨酸tRNA3（tRNASer3）。此外，还讨论了使用Aminex A - 28作为分离tRNA的色谱系统。
