Differential Expression,Cloning and Bioinformatics of FABP2 Gene in Tissues of Fattening Pigs Fed with Different Proportions of Zanthoxylum Seeds

doi:10.7668/hbnxb.20193156

Abstract

Abstract:

The purpose of this study was to analyze the differential expression of FABP2 in different tissues of DLY fattening pigs fed different proportions of Zanthoxylum seeds instead of some corn,and to find the key gene for intramuscular fat deposition;at the same time,the sequence of the coding region of FABP2 in DLY fattening pigs was cloned and analyzed by bioinformatics methods.A total of 288,90-day-old Duroc Landrace Yorkshire fattening pigs(33 kg)were randomly divided into 4 groups with 4 replicates and 18 pigs in each replicate;the control group was fed the basal diet and the test group was fed Zanthoxylum seeds instead of corn in the diet at 2.5%(test group Ⅰ),5.0%(test group Ⅱ)and 7.5%(test group Ⅲ),respectively,for 7 d in the pretest and 100 d in the test period.PCR was used to detect the relative expression of FABP2 in different tissues of the test groups,and the coding region of FABP2 in DLY fattening pigs was also cloned,and bioinformatics analysis software was used to predict the physicochemical properties,structural domains and phosphorylation kinase sites of FABP2 protein.The results showed that FABP2 was differentially expressed in all tissues,with significantly higher expression in jejunum and liver.Compared with the control group,FABP2 was significantly down-regulated in heart,lung,kidney,duodenum,jejunum and cecum tissues in each test group;FABP2 was significantly up-regulated in spleen and ileum tissues in test group Ⅱ and test group Ⅲ;FABP2 was significantly up-regulated in liver,colon,rectum and dorsal muscle tissues in test group Ⅰ,indicating that the addition of Zanthoxylum seeds in the diet had a significant effect on the expression profile of FABP2 gene in DLY fattening pigs.In this experiment,the full length(399 bp)of the coding region of FABP2 was successfully cloned and identified,encoding 132 amino acids,with two missense mutations;FABP2 protein was an acidic stable non-secretory protein with a secondary structure consisting mainly of extended chains and random coils,containing a Lipocalin-FABP2 superfamily conserved structural domain and three disordered regions.

Key words: DLY fattening pigs, Zanthoxylum seeds, FABP2 gene, Tissue expression, Cloning, Bioinformatics analysis

LI Jie, HUANG Xiaoyu, SHANG Xuefeng, YANG Jiaojiao, ZHANG Juanli, XIE Kaihui, ZHANG Yali, YAN Zunqiang, WANG Pengfei, GAO Xiaoli, YANG Qiaoli, MA Yanping, GUN Shuangbao. Differential Expression,Cloning and Bioinformatics of FABP2 Gene in Tissues of Fattening Pigs Fed with Different Proportions of Zanthoxylum Seeds[J]. Acta Agriculturae Boreali-Sinica, 2022, 37(6): 228-238. doi: 10.7668/hbnxb.20193156.

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References 33

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基因名 Gene name	转录本 Transcript No.	引物序列(5'—3') Primer sequences (5'—3')	扩增长度/bp Length	用途 Purpose
FABP2	NM_001031780.1	F: GGTGCCTGGAAGATAGACCG	224	荧光定量
		R: AGTTCAGTTCCGTCTGCGAG
FABP2	NM_001031780.1	F: GCTGGAGACTATGGCATTTGAT	427	PCR扩增
		R: GTGCTCCAAGAACAATGCTCAAT
GAPDH	NM_001206359.1	F: AGTATGATTCCACCCACGGC	139	荧光定量
		R: TACGTAGCACCAGCATCACC

基因名 Gene name	转录本 Transcript No.	引物序列(5'—3') Primer sequences (5'—3')	扩增长度/bp Length	用途 Purpose
FABP2	NM_001031780.1	F: GGTGCCTGGAAGATAGACCG	224	荧光定量
		R: AGTTCAGTTCCGTCTGCGAG
FABP2	NM_001031780.1	F: GCTGGAGACTATGGCATTTGAT	427	PCR扩增
		R: GTGCTCCAAGAACAATGCTCAAT
GAPDH	NM_001206359.1	F: AGTATGATTCCACCCACGGC	139	荧光定量
		R: TACGTAGCACCAGCATCACC

软件Software	功能用途Function	网址Website
Protparam	氨基酸理化性质分析	https://web.expasy.org/protparam/
Sopma	蛋白的二级结构预测	https://npsa-prabi.ibcp.fr/cgi-bin/npsa_automat.pl?page=npsa_sopma.html
Phyre2	蛋白三级结构模型建立	http://www.sbg.bio.ic.ac.uk/phyre2/html/page.cgi?id=index
ExPASy-Protscale	蛋白亲疏水性分析	https://web.expasy.org/protscale/
TMHMM Server.v.2.0	跨膜螺旋结构预测	http://www.cbs.dtu.dk/services/TMHMM-2.0/
PSORT Ⅱ Prediction	蛋白亚细胞定位	https://psort.hgc.jp/form2.html
SignalP 4.1	信号肽裂解位点预测	http://www.cbs.dtu.dk/services/SignalP/
NetPhos 3.1 Server	激酶磷酸化位点分析	http://www.cbs.dtu.dk/services/NetPhos/
NetNGlyc 1.0	N端糖基化定位	http://www.cbs.dtu.dk/services/NetNGlyc/
Conserved domains (CDD)	蛋白保守结构域预测	https://www.ncbi.nlm.nih.gov/cdd
FoldIndex	蛋白折叠无序化分析	http://bioportal.weizmann.ac.il/fldbin/findex

软件Software	功能用途Function	网址Website
Protparam	氨基酸理化性质分析	https://web.expasy.org/protparam/
Sopma	蛋白的二级结构预测	https://npsa-prabi.ibcp.fr/cgi-bin/npsa_automat.pl?page=npsa_sopma.html
Phyre2	蛋白三级结构模型建立	http://www.sbg.bio.ic.ac.uk/phyre2/html/page.cgi?id=index
ExPASy-Protscale	蛋白亲疏水性分析	https://web.expasy.org/protscale/
TMHMM Server.v.2.0	跨膜螺旋结构预测	http://www.cbs.dtu.dk/services/TMHMM-2.0/
PSORT Ⅱ Prediction	蛋白亚细胞定位	https://psort.hgc.jp/form2.html
SignalP 4.1	信号肽裂解位点预测	http://www.cbs.dtu.dk/services/SignalP/
NetPhos 3.1 Server	激酶磷酸化位点分析	http://www.cbs.dtu.dk/services/NetPhos/
NetNGlyc 1.0	N端糖基化定位	http://www.cbs.dtu.dk/services/NetNGlyc/
Conserved domains (CDD)	蛋白保守结构域预测	https://www.ncbi.nlm.nih.gov/cdd
FoldIndex	蛋白折叠无序化分析	http://bioportal.weizmann.ac.il/fldbin/findex

物种Species	1	2	3	4	5	6	7	8	9
三元杂交育肥猪DLY fattening pigs
绵羊Ovis aries	0.13
马Equus caballus	0.25	0.20
牛Bos taurus	0.14	0.01	0.21
犬Canis lupus familiaris	0.11	0.11	0.20	0.11
智人Homo sapiens	0.15	0.15	0.23	0.16	0.14
猕猴Macaca mulatta	0.14	0.14	0.21	0.14	0.13	0.05
小鼠Mus musculus	0.22	0.20	0.30	0.21	0.19	0.22	0.23
大鼠Rattus norvegicus	0.18	0.17	0.27	0.17	0.14	0.18	0.17	0.08
鸡Gallus gallus	0.27	0.27	0.33	0.27	0.23	0.30	0.27	0.22	0.22

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