斯氏副柔线虫GP基因的克隆、抗原表位预测及生物信息学分析

doi:10.16431/j.cnki.1671-7236.2018.06.028

Abstract

Abstract:

This study was aimed to clone and carry out bioinformatics analysis on the sequence of glycoprotein (GP) gene of Parabronema skrjabini.RNA was extracted from Parabronema skrjabini and cDNA was obtained by reverse transcription.A pair of specific primers was designed to amplify the coding region of GP gene.The target gene was linked into cloning vector pMD19-T to be sequenced.The antigen epitopes,physical and chemical properties,signal peptide and transmembrane domain of GP gene encoding protein were predicted according to the bioinformatics analysis.The bioinformatic analysis results showed that GP gene contained a 1 149 bp open reading fram (ORF),which encoding a hydrophobic protein 382 amino acids with a predicted theoretical molecular weight of 40.15 ku and the isoelectric point (pI) of 4.37.Amino acid sequence analysis showed that the GP protein was hydrophobin and unstable whose formula was C₁₇₆₀H₂₈₇₈N₄₅₈O₅₉₀S₁₇₆₀,the grand average of hydropathicity (GRAVY) was 0.032,and the coefficient of instability was 42.43.The protein was absence of signal peptide cleavage sites and the phosporylation site located in serine,threonine and tyrsine residue.The secondary structure of GP was mainly composed of alpha-helix and random curl,consistent with the results of tertiary structure prediction.Antigen epitopes prediction showed that GP protein might have six B-cell epitopes and eight T-cell epitopes.The GP antigen had promise as a candidate for immune diagnostics and a vaccine against Parabronema skrjabini.The GP gene of Parabronema skrjabini was cloned successfully and the structure and antigen epitopes of its coding protein were predicted by bioinformation software.This research would provide the theoretical basis for establishment of iELISA diagnostic method and development of DNA vaccine.

Key words: Parabronema skrjabini; GP gene; antigen epitopes; bioinformatics

CLC Number:

S852.73⁺1

ZHAO Xueliang, WANG Mengya, SUN Ke, FENG Chenchen, WANG Wenlong. Cloning, Prediction of Epitopes and Bioinformatic Analysis of GP Gene in Parabronema skrjabini[J]. , 2018, 45(6): 1645-1652.

References

[1] SEYED S H.Molecular characterization of the first internal transcribed spacer of rDNA of Parabronema skrjabini for the first time in sheep[J].Annals of Tropical Medicine and Parasitology,2015,61(4):241-246.
[2] HASHEMINASAB S S,JALOUSIAN F,MESHGI B.Molecular and morphological characterization of Parabronema skrjabini of sheep and goats at three different geographical zones in Iran[J].Annals of Tropical Medicine and Parasitology,2016,62(1):55-61.(in Chinese)
[3] 赵治国,王海艳,郭铁筝,等.阿拉善双峰驼斯氏副柔线虫感染情况及病理组织学变化[J].动物医学进展,2012,33(11):56-58. ZHAO Z G,WANG H Y,GUO T Z,et al.Infection situation of Parabronema skrjabini and pathological lesions in Alashan bactrian camels[J].Progress in Veterinary Medicine,2012,33(11):56-58.(in Chinese)
[4] 杨莲茹,杨晓野,刘珍莲,等.内蒙古地区骆驼斯氏副柔线虫(Parabronema skrjabini)病调查[J].内蒙古农业大学学报(自然科学版),2004,25(1):43-45. YANG L R,YANG X Y,LIU Z L,et al.Investigation of Parabronema skrjabini disease of camels in Inner Mongolia region[J].Journal of Inner Mongolia Agricultural University,2004,25(1):43-45.(in Chinese)
[5] SONG J,HE Q F.Bioinformatics analysis of the structure and linear B-cell epitopes of aquaporin-3 from Schistosoma japonicum[J].Asian Pacific Journal of Tropical Medicine,2012,5(2):107-109.
[6] 冯陈晨,王文龙,呼和巴特尔.骆驼斯氏副柔线虫转录组的高通量测序及分析[J].中国兽医学报,2017,37(4):671-675. FENG C C,WANG W L,HUHE B.Sequencing and analysis of the transcriptome of Parabronema skrjabini in camel[J]. Chinese Journal of Veterinary Science,2017,37(4):671-675.(in Chinese)
[7] HU R L,ZHANG S F,FOOKS A R,et al.Prevention of rabies virus infection in dogs by a recombinant canine adenovirus type-2 encoding the rabies virus glycoprotein[J].Microbes and Infection,2006,8(4):1090-1097.
[8] NOGAROL C,DECARO N,BERTOLOTTI L,et al.Pestivirus infection in cattle dairy farms:E2 glycoprotein ELISA reveals the presence of bovine viral diarrhea virus type 2 in northwestern Italy[J].Bmc Veterinary Research,2017,13(1):377-386.
[9] STARODUBOVA E S,KUZMENKO Y V,LATANOVA A A,et al.Creation of DNA vaccine vector based on codon-optimized gene of rabies virus glycoprotein (G protein) with consensus amino acid sequence[J].Molecular Biology,2016,50(2):376-380.
[10] CHEN Y J,YU P,LUO J C,et al.Secreted protein prediction system combining CJ-SPHMM,TMHMM,and PSORT[J].Mammalian Genome,2003,14(12):859-865.
[11] PETERSEN T N,BRUNAK S,VON H G,et al.SignalP 4.0:Discriminating signal peptides from transmembrane regions[J].Nature Methods,2011,8(10):785-786.
[12] BIASINI M,BIENERT S,WATERHOUS A,et al.SWISS-MODEL modelling protein tertiary and quaternary structure using evolutionary information[J].Nucleic Acids Research,2014,42(W1):252-258.
[13] FERRET Y,CAILLAULT A,SEBDA S,et al.Multi-loci diagnosis of acute lymphoblastic leukaemia with high-throughput sequencing and bioinformatics analysis[J].British Journal of Haematology,2016,173(3):413-420.
[14] BRITTON C,ROBERTS B,MARKS N D.Functional genomics tools for Haemonchus contortus and lessons from other helminths[J]. Advances in Parasitology,2016,93:599-623.
[15] 张宇飞,李晓霞,王勇士,等.北极狐及乌苏里貉抑制素α亚基基因克隆及生物信息学分析[J].中国畜牧兽医,2017,44(12):3391-3400. ZHANG Y F,LI X X,WANG Y S,et al.Cloning and bioinformatics analysis of inhibin α-subunit gene of arctic fox and Wusuli raccoon dog[J].China Animal Husbandry & Veterinary Medicine,2017,44(12):3391-3400.(in Chinese)
[16] 朱事康,佟铁铸,刘星,等.猪嗜淋巴细胞疱疹病毒2型GD27株gB基因生物信息学分析[J].中国畜牧兽医,2017,44(1):186-193. ZHU S K,TONG T Z,LIU X,et al.Bioinformatics analysis of gB gene of porcine lymphotropic herpesviruses strain GD27[J].China Animal Husbandry & Veterinary Medicine,2017,44(1):186-193.(in Chinese)
[17] TRITTEN L,CLARKE D,TIMMINS S,et al.Dirofilaria immitis exhibits sex-and stage-specific differences in excretory/secretory miRNA and protein profiles[J].Veterinary Parasitology,2016,232:1-7.
[18] DJAFSIA B,NDJOKA D,DIKIT J V,et al.Immune recognition of excretory and secretory products of the filarial nematode Onchocerca ochengi in cattle and human sera[J].Journal of Helminthology,2015,11:1-9.
[19] 李静,刘建华,付强,等.马疱疹病毒1型gB糖蛋白线性B细胞表位的预测与鉴定[J].中国畜牧兽医,2017,44(5):1484-1492. LI J,LIU J H,FU Q,et al.Prediction and identification of linear B-cell epitopes in gB glycoprotein of equine herpesvirus type 1[J].China Animal Husbandry & Veterinary Medicine,2017,44(5):1484-1490.(in Chinese)
[20] DONG L L,TANG R,ZHAI Y J,et al.DNA vaccine expressing herpes simplex virus 1 glycoprotein C and D protects mice against herpes simplex keratitis[J].International Journal of Ophthalmology,2017,10(11):1633-1639.
[21] 张雅岭,王凯,陈芳玲,等.伊氏锥虫变异表面糖蛋白抗原双抗体夹心ELISA检测方法的建立及初步应用[J].中国畜牧兽医,2014,41(2):6-10. ZHANG Y L,WANG K,CHEN F L,et al.Development and preliminary application of double antibody sandwich ELISA for detection of Trypanosoma evansi VSG antigen[J].China Animal Husbandry & Veterinary Medicine,2014,41(2):6-10.(in Chinese)
[22] 马凡舒,张蕾,王洋,等.B细胞抗原表位预测方法的研究进展[J].中国畜牧兽医,2016,43(1):63-67. MA F S,ZHANG L,WANG Y,et al.Advance on prediction methods of B-cell antigen epitope[J].China Animal Husbandry & Veterinary Medicine,2016,43(1):63-67.(in Chinese)

Cloning, Prediction of Epitopes and Bioinformatic Analysis of GP Gene in Parabronema skrjabini

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	WANG Saiqiao, ZHAO Lu, ZHAI Zhenhan, ZHANG Binglei, JIA Wanhang, WANG Yuqin. Tissue Expression and Bioinformatics Analysis of miR-449a/b Precursor Sequences in Sheep [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2175-2184.
[2]	WEI Mingbang, BIANBA Qiongda, XIAO Qingqing, DUAN Mengqi, CHAMBA Yangzom, SHANG Peng. Cloning,Bioinformatics and Expression Analysis of CDKN1B Gene in Tibetan Pigs [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2196-2206.
[3]	LI Yongbin, YANG Huilin, SONG Zihao, ZHANG Yu, YANG Jin, MENG Jia, CUI Xiaozhen, LYU Xiaoling, ZHENG Mingxue, BAI Rui. Cloning and Bioinformatics Analysis of MIF Gene in Eimeria tenella [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2450-2459.
[4]	ZHANG Dianqi, MA Xinhao, YANG Zhimei, MEI Chugang, ZAN Linsen. Bioinformatics Analysis of PAI1 Gene and Its Effect on the Proliferation and Differentiation of Intramuscular Adipocytes in Qinchuan Beef Cattle [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 1729-1741.
[5]	WANG Haorui, YU Baojun, YANG Jiayi, WEI Rong, FU Xi, WANG Chuanchuan, ZHANG Di, LI Desheng, CAI Zhengyun, GU Yaling, ZHANG Juan. Bioinformatics Analysis and Tissue Expression of DERA Gene in Jingyuan Chickens [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 1764-1773.
[6]	CHEN Meiting, ZHENG Congsen, LIANG Zexian, LIN Qiaoer, CHANG Chuanzhe, ZHOU Jun, FENG Jun, LI Linlin, QIN Limei. Cloning and Bioinformatics Analysis of env Gene of Avian Leukosis Virus Subgroup J Strain [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 1785-1795.
[7]	PAN Pengcheng, LEI Zongquan, LI Xian, HU Xiangyun, QIN Qiantao, QIN Zhaoxian, GUAN Zhihui, CHEN Baojian, XIE Bingkun. Bioinformatics Analysis,Eukaryotic Expression Vector Construction and Tissue Expression of MYL2 Gene in Luchuan Pigs [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 1796-1806.
[8]	LIU Jialong, ZHANG Yiwen, ZHANG Yueran, SUN Qijuan, CHEN Jian, HE Lei, JIA Yanyan, DING Ke, YU Zuhua. Cloning,Prokaryotic Expression and Bioinformatics Analysis of ELAVL1 Gene in Chicken [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 1807-1817.
[9]	ZHUO Na, ZHU Zhongwu, LI Jing, HU Shixiang, WANG Jianchang, LIN Hua, CHEN Chaolin, HAN Diangang, AI Jun, CHEN Peifu. Bioinformatics Analysis and Eukaryotic Expression of ORF2 Protein of Swine Hepatitis E Virus [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 1959-1970.
[10]	TAO Qinghua, HUANG Caiyun, CHEN Chao, LIU Wei, SUN Ningning, LI Ang. Cloning and Tissue Expression Difference Analysis of LIF Gene in White Muscovy Duck [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1296-1306.
[11]	FAN Bingfeng, LI Wen, LIU Lixiang, ZHAO Xiangyuan, SHAO Jing, LIN Lefeng, SUN Huimin, ZHANG Xulin, XU Baozeng. Cloning and Bioinformatics Analysis of AANAT Gene in Mink [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1307-1318.
[12]	JIA Xinyue, MA Jing, ZHANG Yanyan, MA Xun, WANG Zhengrong, BO Xinwen. Bioinformatics Analysis and Prokaryotic Expression of EGR-07243 and EGR-07243 Genes of Echinococcus granulosus [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 847-858.
[13]	LI Dan, ZHANG Haisen, WANG Yiqun, GAO Dengke, ZHAO Hongcong, LI Chao, JIN Yaping, CHEN Huatao. Eukaryotic Expression Vector Construction,Bioinformatics and Tissue Expression Profiles Analysis of CRY1 Gene in Dairy Cows [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 870-881.
[14]	SONG Xingchao, MENG Jinzhu, ZHAO Yuanyuan, WU Zhenyang, AN Qingming. Cloning and Bioinformatics Analysis of MyoG Gene Promoter Region Sequence in Guizhou White Goat [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 893-903.
[15]	ZHU Xihui, LI Yanlan, WANG Zhiwei, PANG Zheyu, LI Yubao, LIU Cheng, SI Zhenshu, LU Jianbiao, CAO Shengliang, XUE Xijuan. Whole Genome Sequencing of a Resistant Riemerella anatipestifer Strain and Analysis of Antibiotic Resistance and Genetic Evolution [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1140-1149.