Molecular cloning of a novel cDNA from Mus muscular BALB/c mice encoding glycosyl hydrolase family 1: a homolog of human lactase-phlorizin hydrolase.
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 (d). Gene-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 bioinformatics revealed that this cDNA shared extensive sequence homology with human LPH containing a conserved glycosyl hydrolase family 1 motif important for regulating lactase intolerance.


CONCLUSIONS
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.
He
Wei
W
Department of Nutrition and Food Hygiene, Huaxi School of Public Health, Sichuan University, Chengdu 610041, Sichuan, China.
Ji
Zhen-Yu
ZY
Huang
Cheng-Yu
CY
D000818:Animals; D003001:Cloning, Molecular; D018076:DNA, Complementary; D006801:Humans; D010694:Lactase-Phlorizin Hydrolase / Q000235:genetics*; D051379:Mice; D008807:Mice, Inbred BALB C
D018076:DNA, Complementary; D010694:Lactase-Phlorizin Hydrolase