恶性疟原虫染色体基因文库的构建

THE ESTABLISHMENT OF GENOMIC DNA LIBRARIES FOR THE HUMAN MALARIA PARASITE PLASMOD1VM FALC1PARVM

恶性疟原虫FCC1/HN株DNA经Bam HI部分消化后,取17-22kb片段插入到载体EMBL_4DNA左右臂之间,经体外包装后,感染P_2溶源菌E.coli L_(95),建成基因文库,使原虫基因组中的每一段DNA序列均有99％的几率克隆到文库中。通过原位杂交,筛出了含有重复序列的特异克隆,证明该库稳定而有效。

The DNA of Plasmodium falciparum has been purified and fragmented with restriction endonuclease BamHI. The fragments have been incorporated in vitro into derivatives of bac-teriophage lambda EMBL4 digested with BamHI and Sal I. The recombinant mixture has been ligated and packaged in vitro. The recombinant phages have been identified in E. coli L95host cell and the libraries have been established in which most of the parasite DNA is represented. The ligation proportion of vector to insert is 3 : 1. The recombinant phages of 4×105 have been obtained. By plaque hybridization, we have been able to recover from these libraries specific clones containing repetitive DNA sequences.