Takara DNA vectors are used for cloning as follows.
pUC118/119 DNA
pUC118/119 are plasmid vectors developed for the preparation of single stranded DNA. pUC118 was constructed by inserting intergenic region (IG region) of the M13 phage DNA into the pUC18 plasmid. Therefore, infection by the helper phage M13K07 induces the production of pUC118 as single stranded DNA which is predominantly packaged into phage particles and is then released from bacterial cells. In addition, there is almost no contamination by the single stranded DNA of the helper phage. Using this system, single stranded DNA from large DNA fragments (up to 7 kb) can be stably obtained without deletion.
pTV118N DNA
pTV118N DNA is plasmid vector (phagemids vector) constructed by modifying pUC118 for the preparation of single stranded DNA like pUC118. This plasmid codes lacZ a peptide and is constructed by inserting Nco I cleavage sequence (CCATGG) into the initiation codon (ATG) site of pCU118 plasmid. Therefore, this plasmid induces the expression of foreign gene which is inserted into the Nco I site by utilizing lac promoter, lacZ SD sequence, and the initiation codon and by adjusting the translation frame. The number of bases between SD sequence and the initiation codon is 8, which allows high level expression. In addition, recovery of single-stranded DNA by using helper phage and the subsequent sequencing with RV-N primer enables to read the sequence from the initiation codon site and the translation frame can be confirmed easily.
pKF 18k-2/19k-2 DNA
This DNA is a pUC type vector possessing duplicated amber mutants on Kanamycin resistant gene. When transformed with this DNA, transformants of supE strains like JM109 can grow on plate containing Kanamycin, but transformants of sup 0 strains like MV1184 cannot grow. This DNA is available to site-directed mutagenesis based on ODA method using this property. Moreover, this DNA is available to cloning of a target gene and its expression using lac promoter when used on supE hosts like JM109, like as multi-cloning site as well as pUC18 /19 on lac Z gene, also Nde I restriction site (CATATG) at the position of initiation codon (ATG). It is possible to increase initiation efficiency of translation by using this Nde I site.
