RGD Reference Report - Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology - Rat Genome Database

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Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology

Authors: An, JZ  Song, ZS  Dou, KF  Li, KZ  Han, H 
Citation: An JZ, etal., Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2005 May;21(3):284-6.
RGD ID: 2306976
Pubmed: PMID:15862140   (View Abstract at PubMed)

AIM: To obtain recombinant rat betacellulin with biological activity. METHODS: A 534 bp of rat betacellulin gene fragment was amplified by RT-PCR from rat kidney and cloned into pET28a(+) vector to construct recombinant plasmid pET28a-rBTC. The recombinant plasmid was transformed into E. coli BL-21(DE3) and the betacellulin was expressed under IPTG induction. The expressed betacellulin was detected by SDS-PAGE and Western blot. The expressed protein was purified by Ni(2+) affinity chromatography and then renatured by dialysis. The effect of the renatured protein on proliferation of NIH3T3 cells was detected by MTT colorimetry. RESULTS: Rat betacellulin protein with M(r) being 20 000 was expressed under IPTG induction. The purity of purified protein reached over 96%. After renaturation, the expressed protein could significantly stimulate the proliferation of NIH3T3 cells. CONCLUSION: Rat betacellulin gene is successfully cloned into the expression vector pET28a(+) and highly expressed in E.coli. Purified and refolded betacellulin protein can obviously stimulate the proliferation of NIH3T3 cells.



Gene Ontology Annotations    Click to see Annotation Detail View

Biological Process

  
Object SymbolSpeciesTermQualifierEvidenceWithNotesSourceOriginal Reference(s)
BtcRatpositive regulation of fibroblast proliferation  IDA  RGD 

Objects Annotated

Genes (Rattus norvegicus)
Btc  (betacellulin)


Additional Information