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利用ＲＴ－ＰＣＲ从人肥大前列腺组织钓取９４个氨基酸的人前列腺分泌蛋白（ＰＳＰ９４）全长ｃＤＮＡ，序列分析结果与文献报道的完全一致．将ＰＳＰ９４成熟肽与人ＴＮＦα衍生物（ＴＮＦΔ）通过Ｌｉｎｋｅｒ－ＳＡＰＧＴＰ在基因水平上融合成５′ＰＳＰ９４－ＴＮＦΔ，融合基因ＤＮＡ序列分析结果与设计的相符合．５′ＰＳＰ９４－ＴＮＦΔ在大肠杆菌中表达产物分子量约为３１ｋＤ，表达量约占菌体总蛋白量的３５％．以Ｌ９２９细胞和人前列腺癌细胞株ＰＣ－３为靶细胞进行细胞毒分析结果表明，５′ＰＳＰ９４－ＴＮＦΔ融合蛋白既具有ＴＮＦ的细胞毒活性，又具有对前列腺癌细胞ＰＣ－３的杀伤作用
The full-length cDNA of 94 amino acid human prostatic secretion protein (PSP94) was obtained from human hypertrophy prostatic tissue by RT-PCR, and the sequence analysis results were completely consistent with those reported in the literature. The PSP94 mature peptide and human TNFα derivative (TNFΔ) were fused to 5’PSP94-TNFΔ at the genetic level by Linker-SAPGTP. The DNA sequence analysis of the fusion gene was in accordance with the designed results. 5’PSP94-TNFΔ expressed in E. coli The molecular weight of the product was about 31kD, and the expression amount was about 35% of the total amount of the bacterial cells. Cytotoxicity analysis of L929 cells and human prostate cancer cell line PC-3 showed that the 5’PSP94-TNFΔ fusion protein not only possessed the cytotoxic activity of TNF, but also had a cytotoxic effect on prostate cancer cells PC-3