Yan Z, Xie Y, Li M, Wang P, Wang Y, Bi H, Li Y
Department of Tropical Diseases, Institute of Tropical Medicine, The First Military Medical University, Guangzhou, 510515.
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 1999;17(6):363-6.
To sequence the strong positive clones obtained by immuno-screening of Plasmodium falciparum FCC1/HN lambda gt11 cDNA expression library, and to elucidate the antigenic expressed sequence tags through sequencing the cDNA insert of these positive clones, and new antigenic ESTs could serve as a resource to pursue their corresponding antigen genes.
cDNA inserts of positive lambda gt11 phage clones were amplified by PCR. The PCR products, after purification, were cloned into the M13 mp18 sequencing vector. Single-stranded M13 DNA was prepared and sequenced. Then the acquired sequences were compared in homologies with EMBL/GenBank database on the PC/GENE software system and searched in NCBI (National Center for Biotechnology Information) GenBank using BLAST (Basic Local Alignment Search Tool) commond.
Sequence C03 was part of the known P. falciparum antigenic heat shock protein 70 (Pfhsp70) gene, while the other 5 sequences were new P. falciparum antigenic expressed sequence tags (ESTs).
The 5 new antigenic ESTs generated could serve as the breaking through points in our efforts to find out new P. falciparum antigen genes.