[Construction, expression and identification of recombinant bacillus Calmette-Guérin vaccine secreting human interferon alpha-2a]

[Construction, expression and identification of recombinant bacillus Calmette-Guérin vaccine secreting human interferon alpha-2a] Research Abstract Details 

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[Construction, expression and identification of recombinant bacillus Calmette-Guérin vaccine secreting human interferon alpha-2a] Abstract Text:

Hai-tao Liu,Xiao-wen Sun,Yi-nan Zhang,Jian Zhuo,Jie Fan,Feng Wang,Shu-jie Xia,

OBJECTIVE: To construct a recombinant bacillus Calmette-Guérin vaccine (BCG) secreting human interferon alpha-2a (IFNalpha-2a). METHODS: BCG Ag85B signal sequence was amplified from the genome of BCG by using polymerase chain reaction (PCR) and cloned in E.coli-BCG shuttle-vector pMV261 to get pMS. The cDNA fragment encoding human IFNalpha-2a was amplified from the plasmid pBIFNalpha-2a by using PCR and inserted into the shuttle expression vector pMV261. The recombinant plamid pMSIFNalpha-2a was identified by restriction endonuclease digestion, PCR amplification and nucleotide sequencing. pMSIFNalpha-2a was electroporated into BCG to get rBCG. The DNA and protein expressions of IFNalpha-2a gene in rBCG were determined by PCR and Western blotting respectively. IFNalpha-2a in the culture supernatant of rBCG was detected by enzyme-linked immunosorbent assay (ELISA). RESULTS: The recombinant plamid pMSIFNalpha-2a was constructed successfully and confirmed by restriction endonuclease analysis, PCR detection and nucleotide sequencing analysis. pMSIFNalpha-2a was successfully transformed into BCG by eletroporation and were capable of synthesizing and secreting cytokine IFNalpha-2a. Western blotting revealed that the secretive proteins could specially combine with antibody against human IFNalpha-2a. the level of IFNalpha-2a (324.57 pg/ml) in the culture supernatant of rBCG was higher than control group by ELISA assay. CONCLUSION: The constructed recombinant BCG strain produces and secretes human IFNalpha-2a and it will be used in the treatment of superficial bladder cancer.

[Construction, expression and identification of recombinant bacillus Calmette-Guérin vaccine secreting human interferon alpha-2a] Publishing Authors By Initials

HT Liu,XW Sun,YN Zhang,J Zhuo,J Fan,F Wang,SJ Xia,

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[Construction, expression and identification of recombinant bacillus Calmette-Guérin vaccine secreting human interferon alpha-2a] Journal Published:

PUBLICATION TYPE: Journal Article

Journal: Zhonghua yi xue za zhi

VOLUME: 86

Page Numbers: 2417-20

Journal Abbreviation: Zhonghua Yi Xue Za Zhi

ISSN: 0376-2491

DAY: 12

MONTH: Sep

YEAR: 2006

[Construction, expression and identification of recombinant bacillus Calmette-Guérin vaccine secreting human interferon alpha-2a] Information

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LANGUAGE: chi

NlmUniqueID: 7511141

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Grant and Affiliation Information for [Construction, expression and identification of recombinant bacillus Calmette-Guérin vaccine secreting human interferon alpha-2a]

AFFILIATION: Department of Urology, The First People's Hospital of Shanghai, Jiaotong University, Shanghai, China.

Country: China

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MEDLINETA: Zhonghua Yi Xue Za Zhi

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