#Fugu preparation pdf

The present invention has designed the primer for interferon-gamma albumen coded sequence, after extracting the total RNA of fresh Fugu rubripes (Temmincket Schlegel) kidney with TRIzol Reagent, carry out RT-PCR amplification, RT-PCR product is connected to obtain recombinant plasmid with pMD19T carrier, after double digestion, goal gene is connected with expression vector pET-32a (+) again, obtains recombinant expression plasmid and is converted in e. Descriptionįugu rubripes) in China, be mainly distributed in the Huanghai Sea, the Bohai Sea and the East Sea, it is coastal waters bottom predacious fish, its meat flavour is delicious, there is very high edibleness, in the internal organ such as the sexual gland of Fugu rubripes (Temmincket Schlegel) and liver, produce tetraodotoxin, the biological characteristic of its high reactivity and high specific has potential pharmaceutical developments and is worth, and has broad prospects clinically.Interferon, rabbit (interferon, IFN) is that a class is subject to virus infection at body, or while being subject to the induction such as nucleic acid, bacterial endotoxin, cytokinin, the protein of being secreted by recipient cell, there is broad-spectrum disease resistance cytotoxic activity (being mainly glycoprotein).Research shows that IFN makes cell produce antiviral protein by cell surface receptor effect, suppresses virus copying in host cell.IFN also can strengthen natural killer cell (NK cell) in addition, the lymphocytic vigor of scavenger cell and T, thus play immunoregulation effect, strengthen anti-virus ability, cell growth and cytodifferentiation also have very important adjusting active.Yet from Fugu rubripes (Temmincket Schlegel), directly complicated operation, the cost of this albumen of extraction are high, output is few, can not large-scale industrialized production. The preparation method comprises steps of: extracting the total RNA of fresh liver of fugu rubripes and performing inverse transcription to synthesize cDNA of a first strand performing RT-PCR amplification by adopting the cDNA of the first strand as a template, with the upstream primer being N I:5'-CATGCCATGGGCTCCTACATCCCAGTAGAAATG-3', and with the downstream primer being X I:5'-CCGCTCGAGATGACCTCGAATCGACATCT-3' establishing a clone vector, expressing the vector, transferring into an expression bacterial, and cultivating the bacterial strain to obtain strain subjecting the collected strain to enlarging cultivation, inducing with IPTG and collecting the bacterial strain and smashing the bacterial strain, lysing, purifying, and collecting purified liquid.

The method has advantages of simple operation, low cost, high purity of the recombinant protein and high yield, and is suitable for industrial production. The invention discloses a preparation method of a recombinant fugu rubripe interferon gamma protein. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.) Filing date Publication date Application filed by Dalian Ocean University filed Critical Dalian Ocean University Priority to CN201310640811.3A priority Critical patent/CN103667305A/en Publication of CN103667305A publication Critical patent/CN103667305A/en Status Pending legal-status Critical Current Links

Original Assignee Dalian Ocean University Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)

Inventor 王秀利马普刘洋姜志强仇雪梅 Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.) Pending Application number CN201310640811.3A Other languages Chinese ( zh)

#Fugu preparation pdf

Google Patents Preparation method of recombinant fugu rubripe interferon gamma proteinĭownload PDF Info Publication number CN103667305A CN103667305A CN201310640811.3A CN201310640811A CN103667305A CN 103667305 A CN103667305 A CN 103667305A CN 201310640811 A CN201310640811 A CN 201310640811A CN 103667305 A CN103667305 A CN 103667305A Authority CN China Prior art keywords preparation interferon fugu rubripes bacterial fugu Prior art date Legal status (The legal status is an assumption and is not a legal conclusion. Google Patents CN103667305A - Preparation method of recombinant fugu rubripe interferon gamma protein CN103667305A - Preparation method of recombinant fugu rubripe interferon gamma protein