purified as discussed no above the annealed oligonucleotides were mixed and ligated together in three separate ligation steps shown in the center of fig2 using the procedure described above this produced full length dna 1 u03bcg of the full length dna was subjected to pcr with primers seq id no 32 and seq id no 33 as discussed above the primers provide xbai and bamhi restriction sites permitting the gene to be inserted into a pet11d plasmid vector the double stranded full length pcr product namely the gene of the 2325p6 protein was purified from agarose gel and ligated into a pet 11d plasmid vector at xbai and bamhi restriction site all using the procedure discussed above then using the same procedure described above e coli xl1 blue competent cells were transformed and the recombinant plasmid pet11d 2325p6 dna was used to transform the expression host e coli bl21 de3 competent cells to express the target gene the expressed protein has the amino acid sequence shown in seq id nd 60 in which an additional n terminal methionine amino acid is followed by lysine the first amino acid of the 2325p6 protein the n terminal additional methionine residue was cleaved as stated above to yield 2325p6 recombinant protein having the amino acid sequence seq id no 17 as stated in u s pat no 6.