Mouse Pem, a homeobox gene, encodes a protein consisting of 210 amino acid residues. To study the function of mouse Pem protein, we used the yeast two-hybrid system to screen the library of 7-day mouse embryo with full-length mouse Pem cDNA. Fifty-two colonies were obtained after 1.57×108 colonies were screened by nutrition limitation and β-galactosidase assay. Seven individual insert fragments were obtained from the library, and three of them were identified, one of which was confirmed to be the cell division cycle 37 (Cdc37) homolog gene by sequencing. The interaction between mouse Pem and Cdc37 homolog was then confirmed by glutathione S-transferase pull-down assay, and the possible interaction model was suggested.

人巨细胞病毒是一种DNA病毒，在人群中一般呈亚临床感染和潜伏感染。为研究病毒基因沉默工具和抗病毒制剂，以人巨细胞病毒UL54基因mRNA序列设计互补的外部引导序列，共价结合到大肠杆菌来源RNase P催化核心M1 RNA上，从而构建成MIGS-T6核酶。通过对DNA聚合酶UL54基因亚克隆片段转录产物体外切割研究，证实该核酶具备对UL54 mRNA片段的特异切割能力。

干扰素(I型、II型)是具有抗肿瘤作用的蛋白质型的细胞因子，与4一叠氮苯甲酸反应，经红外光谱确认生成物在约2118cm-1处有叠氮基团的典型吸收，证明合成得到了光活性的干扰素。采用光固定法将光活性蛋白质固定到24孔组织培养聚苯乙烯板上，制成生物材料。实验通过扫描电子显微镜(SEM)，对光固定化细胞因子的微观形态进行观察。通过做细胞生长曲线，透射电镜观察两种细胞因子固定化抑制宫颈癌细胞生长、诱导细胞凋亡的情况。实验表明，共固定IFN-α和IFN-γ能有效地抑制宫颈癌细胞的生长，在低剂量（60ng/孔）的情况下，抑制活性达到70%，而IFN-γ具有明显的协同抑癌作用。

Seven sequence-specific ribozymes (M1GS RNAs) derived in vitro from the catalytic RNA subunit of Escherichia coli RNase P and targeting the mRNAs transcribed by the UL54 gene encoding the DNA polymerase of human cytomegalovirus were screened from 11 ribozymes that were designed based on four rules: (1) the NCCA-3' terminal must be unpaired with the substrate; (2) the guide sequence (GS) must be at least 12nt in length; (3) the eighth nucleotide must be U, counting from the site–1; and (4) around the cleavage site, the sites–1/+1/+2 must be U/G/C or C/G/C. Further investigation of the factors affecting thecleavage effect and the optimal ratio for M1GS/substrate was carried out. It was determined that the optimal ratio for M1GS/substrate was 2:1 and too much M1GS led to substrate degrading. As indicated above, several M1GS that cleaved HCMV UL54 RNA segments in vitro were successfully designed and constructed. Our studies support the use of ribozyme M1GS as antisense molecules to silence HCMV mRNA in vitro, and usingthe selection procedure as a general approach for the engineeringof RNase Pribozymes.

为了研究RNA干涉抑制HCMV UU9基因的作用，以pIXSN(U6启动子)为模板通过两步PCR的方法扩增含U6启动子的siRNA表达片段，并通过TA克隆将siRNA表达片段克隆到pMD18-T载体构建成siRNA表达质粒，同时以人巨细胞病毒AD169病毒株基因组为模板PCR扩增UIA9基因，将其克隆到pEGFP-N1构建融合质粒pEGFP-UIA9。通过脂质体介导将siRNA表达质粒和pEGFP-UU9质粒共转染人宫颈癌细胞系HeLa，在荧光显微镜下观察RNA干涉结果。通过这种方法得到具有介导RNA干涉的siRNA片段，为UM9基因沉默研究提供技术基础。