Objective To study the mechanism of lactose intolerance （LI） by cloning the mouse lactase cDNA and recombining a vector. Methods Total murine RNA was isolated from the small intestine of a 4-week-old BALB/c mouse ...Objective To study the mechanism of lactose intolerance （LI） by cloning the mouse lactase cDNA and recombining a vector. Methods Total murine RNA was isolated from the small intestine of a 4-week-old BALB/c mouse （δ）. Crene-specific primers were designed and synthesized according to the cDNA sequences of lactase-phlorizin hydrolase （LPH） in human, rat, and rabbit. A coding sequence （CDS） fragment was obtained using RT-PCR, and inserted into a clone vector pNEB-193, then the cDNA was sequenced and analyzed using bioinformatics. Results The cDNA from the BALB/c mouse with 912 bp encoding 303 amino acid residues. Analysis of the deduced amino acid sequence using bioinforrnatics revealed that this cDNA shared extensive sequence homology with human LPH containing a conserved glycosyl hydrolase family 1 motif important for regulating lactase intolerance. Conclusion BALB/c mouse LPH cDNA （GenBank accession No： AY751548） provides a necessary foundation for study of the biological function and regulatory mechanism of the lactose intolerance in mice.展开更多
基金This work was supported by National Natural Science Foundation China (No. 30271126).
文摘Objective To study the mechanism of lactose intolerance （LI） by cloning the mouse lactase cDNA and recombining a vector. Methods Total murine RNA was isolated from the small intestine of a 4-week-old BALB/c mouse （δ）. Crene-specific primers were designed and synthesized according to the cDNA sequences of lactase-phlorizin hydrolase （LPH） in human, rat, and rabbit. A coding sequence （CDS） fragment was obtained using RT-PCR, and inserted into a clone vector pNEB-193, then the cDNA was sequenced and analyzed using bioinformatics. Results The cDNA from the BALB/c mouse with 912 bp encoding 303 amino acid residues. Analysis of the deduced amino acid sequence using bioinforrnatics revealed that this cDNA shared extensive sequence homology with human LPH containing a conserved glycosyl hydrolase family 1 motif important for regulating lactase intolerance. Conclusion BALB/c mouse LPH cDNA （GenBank accession No： AY751548） provides a necessary foundation for study of the biological function and regulatory mechanism of the lactose intolerance in mice.
