Journal of Animal and Plant Sciences
J. Anim. Plant Sci. [ISSN 2071 - 7024]
Volume 18(1): 2668 - 2681. Published May, 2013.
Cloning, sequence identification and tissue transcription profile analysis of novel inwardly rectifying potassium channel KCNJ12 gene from the Chinese water buffalo (Bubalus bubalis)
Jin-long
HUO1*, Wei-zhen LI1*, Yong-yun ZHANG2*, Qiao-ling ZHANG3*, Hai-long
HUO4, Li-xian LIU4, Jing LENG1, Da-lin LI5, Yue-yun YUAN5, Feng YUAN1,
Lian-jun LI1, Yong-wang MIAO1†
1 Faculty of Animal Science and Technology, Yunnan Agricultural University, Kunming 650201, Yunnan, China
2 Teaching Demonstration Center of the Basic Experiments of
Agricultural Majors, Yunnan Agricultural University, Kunming 650201,
Yunnan, China
3 College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, Jilin, China
4 Department of Husbandry and Veterinary, Yunnan Vocational and Technical College of Agriculture, Kunming 650031, Yunnan, China
5 Domestic Animal Breeding and Crossbreed-improvement Station of Yunnan Province, Kunming 650021, Yunnan, China
* These authors contributed equally to this work
† Corresponding author. E-mail: yongwangmiao999@163.com & miaoyw1@ynau.edu.cn
Key words: Water Buffalo; KCNJ12 gene; tissue transcription profile; bioinformatics analysis
1 SUMMARY
Potassium inwardly-rectifying channel, subfamily J, member
12 (KCNJ12) plays a critical role in central ventilator
chemosensitivity, and this role could possibly exhibit developmental
changes. In the present study, water buffalo KCNJ12 cDNA was cloned and
characterized. This novel gene was then deposited into NCBI database
and assigned to accession number KC011846 (amino acids accession number
AGC91909). The complete open reading frame of water buffalo KCNJ12 gene
was 1281 bp encoding a KCNJ12 protein of 427 amino acids with a
molecular weight of 48.44 kDa and a pI of 5.55. The putative proteins
of KCNJ12, which are located in the cytoplasm (94.1%), contain a
conserved domain of Ion_trans_2 superfamily and three potential
transmembrane regions without N-terminal signal peptide, which
indicates that KCNJ12 was non-secretory and membrane-embedded proteins.
Similarity comparisons for amino acid sequences reveal that the water
buffalo KCNJ12 protein shares 99.8%, 99.8%, 96.5%, 96.3%, 94.4% and
94.1% identity with that of Bos taurus, Ovis aries, Pan troglodytes,
Homo sapiens, Rattus norvegicus and Mus musculus. The phylogenetic tree
analysis revealed that the buffalo KCNJ12 gene has a closer genetic
relationship with the KCNJ12 genes of Bos taurus and Ovis aries than
with those of Pan troglodytes, Homo sapiens, Rattus norvegicus and Mus
musculus. Real-time PCR analysis shows that water buffalo KCNJ12 gene
is expressed in various tissues, but at different levels. The
expression levels of this gene are high in longissimus dorsi, fat,
cerebrum and heart, moderate in uterine wall, skin, lung, duodenum,
oviduct, placenta, spleen and mammary gland, non-expressed in liver,
stomach, pancreas and ovary. These data provide a foundation for
further insight into this buffalo gene.
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