Comparison of two gel filtration chromatographic methods for the purification of Lily symptomless virus

百合 色谱法 大小排阻色谱法 离心 生物 灯泡 凝胶电泳 污渍 质谱法 聚丙烯酰胺凝胶电泳 差速离心 分子生物学 化学 生物化学 植物 基因
作者
Guang-Hui Dong,Jianhui Wang,Juan Li,Yun Wang,Zhongkui Xie,Lizhe An
出处
期刊:Journal of Virological Methods [Elsevier BV]
卷期号:139 (2): 125-131 被引量:20
标识
DOI:10.1016/j.jviromet.2006.09.008
摘要

Lily symptomless virus (LSV) occurs frequently in many Lilium species worldwide and often causes developmental abnormalities such as a smaller flower and lower bulb yield. In this study, two moderate and efficient gel filtration chromatography (GFC) methods was compared, these two techniques were, respectively, based on Superdex-200 HR and Sephacryl S-1000 SF. The products purified by the two methods were then characterized by measurements with UV-spectrophotometer, reverse transcriptase (RT)-PCR, transmission electron microscope (TEM), polyacrylamide gel electrophoresis (PAGE), Western blotting and matrix assisted laser desorption-ionisation/time-of-flight mass spectrometry (MALDI-TOF-MS). The final yield of purified LSV by the Superdex-200 HR GFC was 9.4 mg from 50 g of fresh infected tissues of Lanzhou lily. However, from the same amount tissues, only 5.6 mg of LSV were obtained by using Sephacryl S-1000 SF GFC. The Superdex-200 HR method was thus shown to be more suitable for the purification of LSV than the Sephacryl S-1000 SF GFC. The Superdex-200 HR method does not require costly equipment for density centrifugation and ultracentrifugation. Furthermore, it can provide an economical and efficient way to obtain purified products for the preparation of antibodies for serological diagnosis or LSV infection and related investigations.

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