Construction of eukaryotic expression plasmid pSecTag2B-msBlyS expressing mouse soluble B lymphocyte stimulator / 华西口腔医学杂志
West China Journal of Stomatology
;
(6): 145-148, 2004.
Article
in Chinese
| WPRIM
| ID: wpr-319034
ABSTRACT
<p><b>OBJECTIVE</b>The purpose of this study was to clone the soluble form of the mouse BlyS (msBlyS) and insert it into a eukaryotic expression vector pSecTag2B in order to further elucidat the antitumor activity induced by msBlyS expressed by the recombined plasmid pSecTag2B-msBlyS.</p><p><b>METHODS</b>Full length cDNA of mouse soluble BlyS (msBlyS) was amplified by reverse transcription-PCR from total RNA of mouse spleen. The PCR product was ligated directly with linearized vector pCR2.1 supplied in the TA cloning kit. The recombined plasmid pCR2.1-msBlyS which was selected and identified using blue-white screening method and restriction map analysis and the purified original plasmid pSecTag2B were both cut by HindIII and EcoR I. The digested fragments were extracted and purified from low-melting temperature agarose and ligated by T4DNA ligase. The recombined plasmid pSecTag2B-msBlyS were isolated and identified by restricted endonuclease cutting and Sanger dideoxy DNA sequencing.</p><p><b>RESULTS</b>The sequencing data indicated that inserted msBlyS gene had correct DNA sequence and orientation.</p><p><b>CONCLUSION</b>Eukaryotic expression vector pSecTag2B. Expressing mouse BlyS have successfully been cloned. This will provide us an opportunity to do further research work on BlyS.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Plasmids
/
Recombination, Genetic
/
Spleen
/
Polymerase Chain Reaction
/
Tumor Necrosis Factor-alpha
/
Cloning, Molecular
/
Sequence Analysis, DNA
/
Receptors, Tumor Necrosis Factor
/
Epitopes, B-Lymphocyte
/
Cell Biology
Type of study:
Prognostic study
Limits:
Animals
Language:
Chinese
Journal:
West China Journal of Stomatology
Year:
2004
Type:
Article
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