[Effects of trichostatin A on the interaction between HBx and histone deacetylase protein 1].

Objectives: To study the effects of trichostatin A (TSA) on protein-protein interaction between HBx and histone deacetylase protein 1 (HDAC1).

Methods: Both HBx and HDAC1 expressing vectors were constructed by the method of routine molecular cloning. The expression of HBx and HDAC1 were observed by Western blot assay.

The antibody preparation and expression of human Pescadillo.

To explore the biological roles of human Pescadillo and investigate its potential effect on tumorigenesis, the cDNA of Pescadillo was fused with that of GST. After purification and elution, the purified GST-Pescadillo fusion protein was obtained, and the antibody against the fusion protein was generated. Endogenous Pescadillo protein was observed to be remarkably induced by estrogen.

[Preparation of antibody against Memo protein and analysis of expression profile of Memo in mouse tissues].

Aim: To prepare and characterize antibody against Memo protein and to detect the tissue distribution of Memo in mice.

Methods: Fusion protein GST-Memo was expressed and purified, and polyclonal antibody against Memo was prepared by immunizing mice. A FLAG-tagged eukaryotic expression vector pcDNA3-FLAG-Memo was constructed.

[An analysis of the features of HBx protein distributed in liver cells and its expression in E. coli].

Objective: To investigate the features of HBx protein distributed in liver cells and its expression in E. coli.

Methods: The expression vectors encoding the full length HBx and its mutants were constructed by the routine molecular cloning method.

[Construction of recombinant plasmid using Neo/E Technology].

A new neo/E counterselection technique was set up using Red recombination, which could be used in constructing recombinant plasmid. Firstly, linear targeting cassettes were amplified by PCR; secondly, two steps of homology recombination occurred in vivo: (1) The neo/E counterselection targeting cassette, consisting of a unique endonuclease recognition site and an antibiotic resistance gene, was introduced into the targeted region. (2) The neo/E cassette was replaced in the second round of recombination by another linear targeting cassettes DNA fragment carrying the targeted gene.

[Induction of protective immunity in rhesus monkey by inoculation with recombinant fusion protein of cholera toxin B subunit-multivalent epitopes of Plasmodium falciparum].

Rhesus monkeys (5 in each group) were inoculated with recombinant fusion protein of cholera toxin B subunit and multi-valent epitopes of Plasmodium falciparum intranasal or intramuscular (i.m.).

[Malignant transformation of NIH3T3 cells induced by ectopic expression of PC-1 gene].

Objective: To establish a mouse fibroblastic cell line stably transfected with PC-1 gene, and using such cell line to investigate tumor development and progression imposed by the ectopic expression of PC-1 gene.

Methods: Eukaryotic expression vector pcDNA3.1(-)/myc-his-pc-1 was transfected into mouse fibroblast cell line NIH3T3 by lipofectamine.

[A novel system for characterization of the transcription activating proteins in mammalian cells].

A system used for detecting the transcriptional activating activity of the function-unknown gene products in mammalian cells was developed. Based on the plasmid pTet-Off and the eukaryotic expressing vector pCDNA3.1B(-)/myc-his, firstly, we constructed a set of recombinant plasmids namely pZHO1 (for cloning into the foreign gene fragment and as a negative control), pZHO2 (as a positive control).

[Expression of PC-1 gene in tumor tissues, cloning and analysis of its putative promoter region].

Background & Objective: PC-1 is a novel gene which is overexpressed in bone metastasis and androgen independent prostate cancer cell line C4-2. The objective of this study was to evaluate the expression level of PC-1 gene in multiple human tumor and normal tissues, and clone a series of putative PC-1 gene promoter regions and analyze their promoter activities.

Methods: PC-1 gene specific DNA sequence was used as probe to hybridize with total RNA from 10 pairs of tumor and normal tissues; The C4-2 cell genomic DNA was used as a template in the polymerase chain reaction to amplify PC-1 upstream regions from the translation initiation codon.