摘要
本文报道用固相亚磷酰胺法合成人恶性疟杂合多肽抗原基因。基因全长为216bp,分为10个寡聚核苷酸片段分别合成,然后经T4 DNA连接酶按设计顺序连接成完整的杂合抗原基因,重组到噬茵体M13 mp 18 RF DNA内,转染大肠杆菌JM109。用分子杂交和酶切分析筛选出重组克隆体。经序列分析,证明所合成的人恶性疟杂合多肽抗原基因与所设计完全一致。
Human P. falciparum hybrid peptide antigen gene has been synthesized by the solidphase phosphoramidite method with ABI381A DNA synthesizer. The gene is 216bp in length and divided into 10 fragments to synthesis. All synthetic fragments were annealed and ligated with T4 DNA ligase. The product of synthetic gene was recombinded with phage M13 mp18 vector and transfected to E. coli JM109. The recombinants were screened by spot hybridization with ^(32)P-label synthetic fragment. The sequence of synthetic gene was analysed by dideoxynucleotide chain termination method. The recults showed that synthesized human P. falciparum hybrid antigen gene was identical with the designed one.
出处
《生物化学与生物物理进展》
SCIE
CAS
CSCD
北大核心
1991年第4期294-298,共5页
Progress In Biochemistry and Biophysics
关键词
疟疾
恶性
抗原基因
化学合成
克隆
Humans P. falciparum hybrid peptide antigen gene, DNA chemical synthesis, cloning
