Molecular cloning of a novel cDNA from Mus muscular BALB/c mice encoding glycosyl hydrolase family 1: a homolog of human lactase-phlorizin hydrolase.
- Author:
Wei HE
1
;
Zhen-Yu JI
;
Cheng-Yu HUANG
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Cloning, Molecular; DNA, Complementary; Humans; Lactase-Phlorizin Hydrolase; genetics; Mice; Mice, Inbred BALB C
- From: Biomedical and Environmental Sciences 2006;19(5):340-345
- CountryChina
- Language:English
-
Abstract:
OBJECTIVETo 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.
RESULTSThe 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.
CONCLUSIONBALB/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.
