Method of preparing chicken Akirin2 protein through insect cell expression system

• Main Authors: REN GUANGCAI, LIU CHUANGAO, CHEN RUIAI, HUANG MIAORONG
• Format: Patent
• Language: Chinese; English
• Published: 15.06.2016
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS THEREOF; CULTURE MEDIA; ENZYMOLOGY; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

The invention discloses a method of preparing chicken Akirin2 protein through an insect cell expression system. The method includes the steps of: 1) performing codon optimization to the chicken Akirin 2 gene according to insect cell preferred codons; 2) synthesizing an Akirin2 sequence; 3) PCR-amplifying the Akirin2; 4) constructing a baculovirus expression carrier pFastBac1-his-thrombin-Akirin2; 5) performing pFastBac1-his-thrombin-Akirin2 baculovirus carrier package in the insect cell sf9; and 6) expressing and purifying the chicken Akirin2 virus protein. The method has high expression level and can be used for producing the protein having proper translation, which is convenient to modify the protein, thereby simplifying cell growth conditions. 本发明公开了利用昆虫细胞表达系统制备Akirin2蛋白的方法，该方法包括根据昆虫细胞偏爱密码子对鸡Akirin2基因进行密码子优化、Akirin2序列合成、PCR扩增Akirin2、构建杆状病毒表达载体pFastBac1-his-thrombin-Akirin2、在昆虫细胞sf9中进行pFastBac1-his-thrombin-Akirin2杆状病毒载体包装、鸡Akirin2病毒蛋白的表达和纯化等步骤。本发明所述的方法具有较高的表达水平，可以生产的带有适当的翻译蛋白便于蛋白的修饰，简化细胞生长条件。