Cloning and tissue expression analysis of the cytochrome P450c17-Ⅰ (CYP17-Ⅰ) gene from mud loach (Misgurnus anguillicaudatus) and large-scale loach (Paramisgurnus dabryanus)

doi:10.3969/j.issn.1004-1524.2016.11.08

›› 2016, Vol. 28 ›› Issue (11): 1853-1861.DOI: 10.3969/j.issn.1004-1524.2016.11.08

• Animal Science • Previous Articles Next Articles

Cloning and tissue expression analysis of the cytochrome P450c17-Ⅰ (CYP17-Ⅰ) gene from mud loach (Misgurnus anguillicaudatus) and large-scale loach (Paramisgurnus dabryanus)

LIU Shi-li^{1, 2}, ZHANG Ai-ju¹, LIAN Qing-ping¹, WU Wei-jun^{3, 4, *}, JIA Yong-yi¹, JIANG Wen-ping¹, LI Fei¹

1. Zhejiang Institute of Freshwater Fisheries, Agriculture Ministry Key Laboratory of Healthy Freshwater Aquaculture / Key Laboratory of Freshwater Aquatic Animal Genetic and Breeding of Zhejiang Province, Huzhou 313001, China;
2. College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China;
3. Fisheries Technology Extension Station, Water Resources Bureau of Qingyuan County, Qingyuan 323800, China;
4. Fisheries Branch, Hydropower Institute of Qingyuan County, Qingyuan 323800, China

Received:2016-03-14 Online:2016-11-15 Published:2016-11-16

Abstract

Abstract: In this study, full-length cDNA sequences of the cytochrome P450c17-Ⅰ (CYP17-Ⅰ) gene were cloned using homologous cloning and 3'- and 5'-rapid amplification of cDNA ends techniques from mud loach (Misgurnus anguillicaudatus) and large-scale loach (Paramisgurnus dabryanus). Their expression in various tissues were then explored using quantitative real-time PCR (qRT-PCR). In mud loach, the cloned sequence was 1 706 bp in length and contained a 1 563 bp open reading frame (ORF) encoding a protein of 520 amino acids. In the large-scale loach, the cloned sequence was 1 763 bp in length and contained a 1 545 bp ORF encoding a protein of 514 amino acids. Both CYP17-Ⅰ proteins had a predicted signal peptide, a transmembrane domain, a conserved protein domain and three conserved functional areas. Identity analysis showed that CYP17-Ⅰ amino acid sequences were 99% identical between the two loaches, and were more than 70% identical to the same proteins from other fish species. Phylogenetic analysis showed that the closest relationship of CYP17-Ⅰ amino acid sequences was between this two loaches, and the phylogenetic relationships among the CYP17-Ⅰ amino acid sequences were in accord with the traditional classification. CYP17-Ⅰ was expressed widely in the intestine, muscle, heart, stomach, liver, testis, ovary and spleen in the two species of loach, with the highest expression levels being observed in the the testis and ovary.

Key words: cytochrome, Misgurnus anguillicaudatus, Paramisgurnus dabryanus, CYP17-Ⅰ

CLC Number:

S917.4

LIU Shi-li, ZHANG Ai-ju, LIAN Qing-ping, WU Wei-jun, JIA Yong-yi, JIANG Wen-ping, LI Fei. Cloning and tissue expression analysis of the cytochrome P450c17-Ⅰ (CYP17-Ⅰ) gene from mud loach (Misgurnus anguillicaudatus) and large-scale loach (Paramisgurnus dabryanus)[J]. , 2016, 28(11): 1853-1861.

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Cloning and tissue expression analysis of the cytochrome P450c17-Ⅰ (CYP17-Ⅰ) gene from mud loach (Misgurnus anguillicaudatus) and large-scale loach (Paramisgurnus dabryanus)

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