id no 15 and seq id no 16 were synthesized and purified as discussed above in the next step shown at the top of fig1 and described in detail above pairs of oligonucleotides were mixed and annealed to form duplex oligonucleotides a1 a2 a3 a4 a5 a6 and a7 these annealed oligonucleotides a1 a2 a3 a4 a5 a6 and a7 were then agarose gel purified as discussed above the annealed and purified oligonucleotides were then mixed and ligated together in three separate ligation steps shown tin the center of fig1 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 3 and seq id no 16 as discussed above the primers provide xbai and bamhi restriction sites permitting the gene to be inserted in a petlld vector the gene of the 2325p4 protein was agarose gel purified as discussed above the purified 2325p4 gene was then digested with xbai and bamhi restriction enzyme and ligated into a petlld plasmid vector as discussed above then as discussed above the ligated reaction mixture was used to transform e coli xl1 blue competent cells and the recombinant plasmid petlld 2325p4 dna was then used to transform the expression host to express the target gene as discussed above the expressed protein has the amino acid sequence shown in seq id no 59 in which an additional n terminal methionine residue is followed by lysine the first amino acid of the 2325p4 protein the n terminal additional methionine residue was cleaved as stated above_to yield 2325p4 recombinant protein having the amino acid sequence seq id no 1 as stated in u s pat no 6 239 257 b1 2325p4.