Construction of cDNA library from NPC tissue and screening of antigenic genes.
To construct cDNA library of nasopharyngeal carcinoma (NPC) and obtain the NPC associated or specific antigens from it, we used a powerful new method to identify the antigens eliciting humoral immune response, which is SEREX (serological identification of antigen by recombinant cDNA expression library). Autologous serum of NPC patient was used to screen the reactive clones in the human NPC tissue cDNA library consisted of 3.64 million recombinants. The 23 exact positive clones were subcloned to monoclonality and the size of cDNA inserts was identified by PCR. Then the nucleotide sequence of cDNA inserts was determined, and the sequence alignments were performed with BLAST software on GenBank database. They represented 16 different antigens. A detailed sequence analysis showed that 10 of 16 genes were high homologous to genes known in GenBank, such as RPL31, S100 A2, MT2A, etc. However, there were also 6 genes with low homology to genes in GenBank. Furthermore, 3 of 6 genes may be novel genes. The associations of these genes to NPC and the roles that they played in the occurrence and development of NPC should be further revealed.
Shu
Jun
J
Cancer Research Institute, XiangYa School of Medicine, Central South University, Changsha, China.
He
Xiao Juan
XJ
Li
Guan Cheng
GC
D000951:Antigens, Neoplasm / Q000235:genetics* / Q000276:immunology; D002277:Carcinoma / Q000235:genetics* / Q000276:immunology; D030561:Databases, Nucleic Acid; D015723:Gene Library; D006801:Humans; D009303:Nasopharyngeal Neoplasms / Q000235:genetics* / Q000276:immunology; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid
D000951:Antigens, Neoplasm