MOLECULAR AND CELLULAR NUTRITION

Cloning, Sequence Analysis and Tissue Expression of Donkey (Equus asinus) Oligopeptide Transporter 1 Gene

  • ZHOU Miaomiao ,
  • LIU Guiqin ,
  • LIU Wenqiang ,
  • ZHU Mingxia ,
  • WANG Changfa
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  • Liaocheng Research Institute of Donkey High-Efficiency Breeding and Ecological Feeding, College of Agriculture, Liaocheng University, Liaocheng 252000, China

Received date: 2020-12-30

  Online published: 2021-07-06

Supported by

 

Abstract

Oligopeptide transporter 1(PepT1) plays an important role in the uptake of small peptides from animal intestine. The aim of this study was to study the molecular cloning, sequence analysis and tissue expression of PepT1 gene in donkey (Equus asinus) (ePepT1). Firstly, total RNA was extracted from donkey intestines, and ePepT1 gene fragment was cloned by PCR, sequenced and sequence analyzed. Then, total RNA was extracted from kidney, spleen, heart, liver, lung, stomach, duodenum, jejunum and ileum of donkey, and the relative expression level of ePepT1 gene was determined by real-time quantitative PCR. The results showed that the cloned ePepT1 cDNA sequence was 2 124 bp at full length encoding a 707 amino acids peptide with molecular of 78.6 ku and theoretical soelectric point (pI) of 7.52. Bioinformatics analysis showed that the deduced peptide sequence possessed all the characteristic features of ePepT1 identified. The ePepT1 had 11 potential transmembrane domains (TMD), with a large extracellular loop between TMD 9 and TMD 10. Five putative extracellular N-glycosylation sites, five putative extracellular protein kinase C (PKC) sites, and three protein kinase A (PKA) sites were identified in the ePepT1 sequence. The study of tissue distribution showed that ePepT1 gene was expressed in all tissues, but the relative expression level was the highest in intestinal tract (duodenum, jejunum and ileum). In conclusion, ePepT1 gene is cloned for the first time in this study, and its tissue expression is studied, which provides a basis for further study on the absorption of small peptide in donkey intestine.

Cite this article

ZHOU Miaomiao , LIU Guiqin , LIU Wenqiang , ZHU Mingxia , WANG Changfa . Cloning, Sequence Analysis and Tissue Expression of Donkey (Equus asinus) Oligopeptide Transporter 1 Gene[J]. Chinese Journal of Animal Nutrition, 2021 , 33(7) : 4030 -4037 . DOI: 10.3969/j.issn.1006-267x.2021.07.044

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