the known bacterial porin structures hvdac1 gene from homo sapiens cloned in a general e coli cloning vector pbs u2212 stratagene inc was obtained from dr michael forte at oregon health sciences university portland oreg the gene was subsequently amplified by high fidelity pcr using the pfu turbo polymerase stratagene inc with ends designed to be compatible with the r sphaeroides heterologous platform vector pmems1 after amplification and restriction enzyme digestion the pcr product carried spei and bamhi cohesive ends and no intrinsic stop codon upon insertion into pmems1 via directional cloning into the spei and bgiii compatible ends translational termination and the affinity tag are by design provided in frame by sequences 3 u2032 to the bgiii site in the platform vector depicted in fig6 the pmems1 is constructed using the broad host range vector prk404 after conjugation of the new construct into r sphaeroides and large scale culture of the resulting transconjugants solubilized cytoplasmic and icm fractions were subjected to imac fig7 shows an sds page gel of proteins that bound to the column fig7 depicts a coomassie blue stained 12 5 percent acrylamide gel showing overexpression of a transmembrane h sapiens voltage dependent anion channel in the intracytoplasmic membrane fraction of rhodobacter lanes 1 and 5 are molecular weight markers lane 2 is a control sample of purified reaction centers showing the l m and h subunits lane 3 is the membrane fraction showing the hvdac lane 4 is the soluble fraction from the same cells proteins in lanes 2 4 were purified by imac described supra the results reveal a prominent u02dc 32 kilo dalton kda band in the icm fraction from cells harboring the hvdac constructs the size of this band.