[Molecular Cloning of Smooth Muscle Calponin h1 in SHeep.].
It was found that the level of Calponin h1 (CaP h1) mRNA was significantly up-regulated by Estrogen in the myometrium of sheep towards the end pregnancy. Although the CaP h1 has been widely used as a reference gene to observe the changes of expression level of other genes, the full-length gene in sheep has not been obtained. With the oligo nucleotide primers according to human, mouse and pig CaP h1 mRNA, the full-length cDNA of CaP h1 was cloned by 5'- and 3'-RACE (Genbank accession number = AY327118). The cDNA was 1499bp in length and contained a complete open reading frame of 891 bp, encoding a protein of 297 amino acid residues. 5'-and 3'-UTR was 79 bp and 529bp, respectively. With PCR-SSP approaches,the genomic DNA of sheep CaP h1 was obtained .It showed that the gene has 7 exons and 6 introns, spanning over 8kb(Genbank accession number of introns : AY771807,AY771808, AY771809, AY771810.) Homologous comparison indicated that the cDNA sequences are highly conserved across the species. The highest homology was found in wild pig (92%), followed by human (88%), rat (81%), mouse (81%) and chicken (79%). The intron sequence and length showed a large variation among species (>50%).
Yue
Zuo-Jun
ZJ
Institute of Genentics and Developmental Biology, Chinese Academy of Sciences, Beijing, 100101, China, Email: zjyue@genetics.ac.cn.
Song
Jun-Fang
JF
Huang
Zhao-Feng
ZF
Liu
Hai-Bo
HB
Zhu
Zhao-Hui
ZH
Zhang
Xiao-Juan
XJ
Zhang
Yi
Y
Tan
Ping-Ping
PP
Ma
Run-Lin
RL
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D009130:Muscle, Smooth; D012756:Sheep / Q000235:genetics
D018076:DNA, Complementary