猪CA5B基因的序列特征和表达分析

doi:10.11843/j.issn.0366-6964.2021.02.005

摘要/Abstract

摘要： 旨在通过分析猪CA5B （线粒体碳酸酐酶VB，carbonic anhydrase 5b， mitochondria；CA5B/CAVB/Car5b）基因的序列特征和时空表达，解析CA5B对猪组织器官发育和代谢的影响。本研究采集6头240日龄贵州猪（公、母各3头）的心、肝、脾、肺、肾、皮下脂肪、背最长肌、睾丸、卵巢，采集通城猪和长白猪27个生长发育时间点（出生前33、40、45、50、55、60、65、70、75、80、85、90、95、100、105 d和出生后0、9、20、30、40、60、80、100、120、140、160、180 d）（每个品种3头）的骨骼肌，提取上述组织器官RNA进行转录组测序，分析CA5B基因的组织表达谱以及骨骼肌27个生长发育时间点的时序表达；基于NCBI数据库中CA5B蛋白序列，进行物种间的保守性分析，构建系统进化树，分析蛋白互作网络；通过Targetscan、PicTar和miRanda等软件预测与CA5B结合的潜在miRNAs，并在猪RNA编辑数据库中分析CA5B基因潜在的RNA编辑位点。结果发现，CA5B基因在猪、人、小鼠、猴、牛、马、山羊、绵羊、狗和猫中高度保守，相对于小鼠而言，猪CA5B蛋白优先与人聚为一类。CA5B蛋白与线粒体跨膜运输蛋白、类肌球蛋白卷曲螺旋蛋白、细胞色素家族等互作；CA5B在猪的脂肪、睾丸以及卵巢中高表达，在骨骼肌生长发育过程中胚胎期的表达高于出生后阶段；靶标预测结果表明，CA5B可能受ssc-miR-22-5p调控（P<0.05）；此外，CA5B还存在3个RNA编辑位点。结果提示，CA5B可能在猪骨骼肌生长发育和能量储存利用中起重要作用。

关键词: 猪, CA5B, 时空表达, 序列特征, miRNA, RNA编辑

Abstract: This study aimed to analyze the effects of CA5B (carbonic anhydrase 5b， mitochondria; CA5B/CAVB/Car5b) on the organ/tissue development and metabolism in pigs through investigating its sequence characteristics and spatial expression patterns. The heart, liver, spleen, lung, kidney, subcutaneous fat, longissimus dorsi, testis and ovary tissues of 3 boars and 3 sows of 240-day-old Guizhou pigs were collected, and the skeletal muscle tissues at 27 growth and development time points (33, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105 days before birth and 0, 9, 20, 30, 40, 60, 80, 100, 120, 140, 160, 180 days after birth) of 3 Tongcheng pigs and 3 Landrace pigs were also collected. We performed transcription sequencing using RNA extracted from the above tissues, and analyzed the expression profile of CA5B gene in 9 tissues and it's temporal expression in skeletal muscle at 27 development time points. Then the conservative analysis among species, construction of phylogenetic tree and protein interaction network analysis of CA5B gene were conducted based on its protein sequence extracted from NCBI database. Finally, three softwares (Targetscan, PicTar, miRanda) were used to predict the potential miRNAs binded to CA5B gene. The RNA editing sites located in CA5B gene were also detected using the RNA editing database. The results showed that CA5B gene was highly conservative expressed in pig, human, mouse, monkey, cattle, horse, goat, sheep, dog and cat, moreover, the pig and human had higher homology than mouse based on the protein sequence of CA5B. Interaction network showed that CA5B protein mainly interacted with mitochondrial transmembrane transport protein, myosin-like coiled-coil protein, cytochrome family and other proteins. The CA5B was highly expressed in fat, testis and ovary, and the expression level in embryonic period was higher than that in postnatal period in skeletal muscle. In addition, target prediction results suggested that CA5B might be regulated by ssc-miR-22-5p (P<0.05), and there were 3 RNA editing sites in CA5B. The results suggested that CA5B potentially play an important role in skeletal muscle growth， development，energy storage and utilization in pig.

Key words: pig, CA5B, spatiotemporal expression, sequence characteristics, miRNA, RNA editing

中图分类号:

S828.2

郭晋, 范新浩, 杨亚岚, 梁国明, 唐中林. 猪CA5B基因的序列特征和表达分析[J]. 畜牧兽医学报, 2021, 52(2): 322-330.

GUO Jin, FAN Xinhao, YANG Yalan, LIANG Guoming, TANG Zhonglin. Sequence Characteristics and Expression Analysis of CA5B Gene in Pigs[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(2): 322-330.

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