Publication in refereed journal
香港中文大学研究人员 ( 现职)
| 曾允康先生 (生物医学学院) |
| 韦妙宜教授 (那打素护理学院) |
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摘要The human kiaa0319-like gene is one of the candidate genes for developmental dyslexia, but the exact function of the encoded KIAA0319L (KL) protein is not known. To allow functional analysis a purified, biologically active KL protein is required. The kiaa0319-like gene was expressed in insect cells using the baculovirus expression system. To optimize the expression of the kiaa0319-like gene and to be able to purify the KL protein, several approaches were used. Two different recombinant baculoviruses were made, one with the full length coding sequence of KL and one that lacked the transmembrane domain to facilitate purification. Versions in which the kiaa0319L sequences were cloned downstream of the honeybee melittin signal sequence were also made. All four constructs contained a C-terminal influenza hemagglutinin (HA)-tag. Sf9 insect cells infected with these recombinant baculoviruses produced the KL protein, as demonstrated by Western blot analysis using either the HA-antibody or KL-specific polyclonal serum.
着者Savanne Holster, Monique M. van Oers, Els C. Roode, Otto W. H. Tsang, Venus S. Y. Yeung, Just M. Vlak, Mary M. Y. Waye
期刊名称Journal of Biochemistry and Molecular Biology in the Post Genomic Era
出版年份2012
卷号2
期次1
页次45 - 52
国际标準期刊号2156-5732
语言美式英语
关键词dyslexia, baculovims expression system, kiaa0319, kiaa0319-like
