In order to establish a possible evolutionary correlation between α lactalbumin and lysozyme,site\|directed mutagenesis on α\|lactalbumin cDNA was performed to change α\|lactalbumin into lysozyme.The gene of α\|la...In order to establish a possible evolutionary correlation between α lactalbumin and lysozyme,site\|directed mutagenesis on α\|lactalbumin cDNA was performed to change α\|lactalbumin into lysozyme.The gene of α\|lactalbumin was cloned into phagemid pSK +vector and the single strand template of which was prepared.Thereafter,the mutagenic oligonucleotide primers were designed and used to synthesize the mutated double strand DNA.By means of this site\|directed mutagenesis,the amimo acid residues at position His 32 and Thr 33 of bovine α\|lactalbumin cDNA were substituted with Leu and Glu,respectively.Thus,the catalytic site of lysozyme was obtained in α\|lactalbumin.Furthermore,Tyr 103 was substituted by Ala and the lysozyme substrate binding conformation was formed in α\|lactalbumin as well.The structural as well as functional relationship between α\|lactalbumin and lysozyme indicated that there existed possible evolutionary correlation between the two proteins.展开更多
文摘In order to establish a possible evolutionary correlation between α lactalbumin and lysozyme,site\|directed mutagenesis on α\|lactalbumin cDNA was performed to change α\|lactalbumin into lysozyme.The gene of α\|lactalbumin was cloned into phagemid pSK +vector and the single strand template of which was prepared.Thereafter,the mutagenic oligonucleotide primers were designed and used to synthesize the mutated double strand DNA.By means of this site\|directed mutagenesis,the amimo acid residues at position His 32 and Thr 33 of bovine α\|lactalbumin cDNA were substituted with Leu and Glu,respectively.Thus,the catalytic site of lysozyme was obtained in α\|lactalbumin.Furthermore,Tyr 103 was substituted by Ala and the lysozyme substrate binding conformation was formed in α\|lactalbumin as well.The structural as well as functional relationship between α\|lactalbumin and lysozyme indicated that there existed possible evolutionary correlation between the two proteins.
