Cloning of the mouse SSTR5 gene.
Five somatostatin receptor subtypes (SSTR1-5) have been recently cloned in different mammalian species. To date the mouse SSTR5 gene has not been cloned. Cloning and characterization of these genes allows for molecular analysis of receptor expression and function. Since the mouse has become a mammalian model that is permissive for genetic manipulation, the purpose of this study was to clone and characterize the mouse SSTR5 to investigate the molecular mechanisms of this receptor subtype. The gene was cloned by screening the mouse 129SvJ genomic library with a PCR-amplified probe from the 5' end of the mouse SSTR2 gene. The probe corresponded to the first 210 amino acids spanning from the amino terminal through the fourth transmembrane domain of the SSTR2 receptor. The primary screening generated 22 positive clones. During secondary and tertiary library screening 3 positive clones with the same restriction endonuclease digestive pattern were isolated. The nucleotide sequence of 1 clone was identified using a DNA sequencing technique. The sequence obtained was analyzed using the Basic Local Alignment Search Tool, which performs fast database searching combined with rigorous statistics for judging the significance of matched sequences. The result of this analysis showed that the sequence identified had 92% homology with the rat SSTR5 and 80% homology with the human SSTR5, which suggests that the cloned gene represents the mouse 129SvJ SSTR5 gene. In conclusion the mouse SSTR5 gene was successfully cloned. This gene will be important for use in gene regulation and gene ablation studies to determine the molecular events controlled by SSTR5 in regulating murine physiology and development.
Moldovan
S
S
Department of Surgery, Baylor College of Medicine, Houston, Texas 77030, USA.
DeMayo
F
F
Brunicardi
F C
FC
D000818:Animals; D003001:Cloning, Molecular; D015723:Gene Library; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D051379:Mice; D008969:Molecular Sequence Data; D017481:Receptors, Somatostatin / Q000235:genetics*; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid
D007328:Insulin; D017481:Receptors, Somatostatin; C094573:somatostatin receptor 5