[Steady expression and activity determination of recombinant signal peptide of mBD2 in SiHa cell].

Objective: To observe the expression pattern and effect of recombinant murine beta defensin 2 (rmBD2) on the proliferation of cell transfected with pcDNA3. 1 (+)/rmBD2.

Methods: The recombinant plasmid pcDNA3.

[Cloning of ureI gene from Helicobacter pylori and its expression in E. coli].

Objective: To clone the urea membrane channel gene (ureI) from Helicobacter pylori (Hp) for its expression in E. coli, and evaluate the expression conditions and immunological features of the fusion protein.

Methods: ureI gene cloned by PCR from Hp was inserted into the plasmid pET32a (+) to construct the recombinant plasmid pET32a/ureI, followed by identification by BglII and HindIII digestion and sequencing.

[Cloning and eukaryotic expression of murine beta-defensin-2(mBD-2)].

Aim: To clone murine beta defensin-2 gene (mBD2) and to express the mBD2 protein eukaryotically.

Methods: Total RNA was isolated from the lungs of BALB/c mice which were injected with LPS in advance. The DNA fragment encoding mBD2 was amplified by RT-PCR and inserted into the plasmid pcDNA3.

[Analysis of protein expression feature and construction of procaryotic expression system for human papillomavirus type 16 (HPV-16) E4 gene].

Objective: To clone the E4 gene (E4) from human papillomavirus type 16(HPV-16), construct the engineering bacteria of prokaryotic expression, and explore the expression conditions and the characters of expression product.

Methods: The complete E4 gene was cloned by PCR from the sample cell extract of clinical cervical disease that was the positive HPV-16 confirmed by Real-PCR. The E4 DNA fragment was inserted into the pET32a(+) to construct a prokaryotic expression plasmid, called as pET32/E4.

[Expression and immunogenicity analysis of a recombinant fusion protein of V. Cholera ctB and H. pylori ure I].

Aim: To express fusion protein of the cholera toxin B subunit (ctB) and the urea membrane channel gene (ure I) of H. pylori in E. coli, and analyze its immunogenicity.

[Prokaryotic expression and identification of human papillomavirus type 16 E7 protein encoding gene].

Objective: To construct the prokaryotic expression plasmid pET32/E7 and express the human papillomavirus type 16 E7 protein in E. coli.

Methods: HPV16 E7 gene was amplified by PCR.

[Construction of eukaryotic expressing plasmids for HA and HA1 of influenza A virus and their transient expression in HEK293 cells].

Objective: To construct influenza A virus (A/PR/8/34) HA and HA1 eukaryotic expressing plasmids and study their expression in HEK293 cells.

Methods: HA and HA1 genes were cloned by RT-PCR and then inserted into pcDNA3. 1 (+).

[Prokaryotic expression and identification of human papillomavirus type 16 E5 protein].

Objective: To construct the prokaryotic and eukaryotic expression vectors pET32/E5 and pcDNA3.1/E5 for transformation into E. coli BL21 and NIH(3)T(3) cells respectively to observe the expression of human papillomavirus type 16 E5 protein (HPV16 E5).