Investigation into the LINE transposition process by structural study of DNA-protein complexes

该项目的目的是揭示LINE的结构基础。为了获得稳定的蛋白质，我测试了四个EN-Myb构建体，但没有一个能比Sart 1-ORF 2 p更好地表达或纯化。接下来，我使用Sf 9细胞作为蛋白表达系统来克服密码子使用问题。虽然我检测到ORF 1 p的表达，但ORF 2 p没有共表达。我试图纯化在Sf 9细胞中表达的ORF 1 p，然而，ORF 1 p在不溶性级分中回收并且未能纯化。

The aim of this project is to reveal the structural basis of the LINE. To obtain the stable protein, I tested four EN-Myb constructs, but none could be expressed or purified better than Sart1-ORF2p. Next, I used Sf9 cells as protein expression system to overcome the codon-usage issue. Although I detected the ORF1p expression, ORF2p was not co-expressed. I tried to purify the ORF1p expressed in Sf9 cells, however, ORF1p was recovered in insoluble fraction and failed to purify.