Cloning, expression and functional verification of the serine protease inhibitor gene<em> ApSerpin</em>-<em>FA</em>72 from <em>Anatolica polita borealis</em> (Coleoptera: Tenebrionidae)
昆虫学报
       Home | About Journal | Editorial Board | Subscription | Instruction | Advertisement | Contact Us | Chinese
ACTA ENTOMOLOGICA SINICA  2017, Vol. 60 Issue (8): 857-864    DOI: 10.16380/j.kcxb.2017.08.002
RESEARCH PAPERS Current Issue | Next Issue | Archive | Adv Search Previous Articles  |  Next Articles  
Cloning, expression and functional verification of the serine protease inhibitor gene ApSerpin-FA72 from Anatolica polita borealis (Coleoptera: Tenebrionidae)
ZHAO Yu, LIU Zhong-Yuan*
(Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang University, Urumuqi 830046, China)
 Download: PDF (4513 KB)   HTML (1 KB)   Export: BibTeX | EndNote (RIS)      Supporting Info
Abstract 【Aim】 This study aims to clone and analyze the expression of a serine protease inhibitor gene from Anatolica polita borealis so as to verify its function. 【Methods】 A serine protease inhibitor gene from A. polita borealis was cloned using PCR and RACE methods. Bioinformatics approach was applied to analyze the basic properties of the gene and its coded protein, and to make the multiple sequence alignment with its homologous sequences. The recombinant expression vector pET-32a-ApSerpin-FA72 was constructed. The expression, purification and functional verification of the recombinant target protein were analyzed. 【Results】 A serine protease inhibitor gene was cloned from A. polita borealis and named ApSerpin-FA72 (GenBank accession no.: MF188125). Its opening reading frame (ORF) is 1 176 bp in length, encoding a 391 amino-acid residue with the predicted molecular weight of 43.7 kD and pI of 5.14. ApSerpin-FA72 is a hydrophilic protein with a signal peptide of 21 amino acids at the terminus. It might play roles in the stress response and immune response, and has the highest homology with Tribolium castaneum Serpin. The purified recombinant protein TrxA-ApSerpin-FA72 has the molecular weight of 63.7 kD. The functional verification results revealed that the recombinant protein TrxA-ApSerpin-FA72 had an inhibitory effect on the activities of trypsin and chymotrypsin. 【Conclusion】 The expression product of Anatolica polita serine protease inhibitor gene has an inhibitory effect on trypsin and chymotrypsin, indicating that it may inhibit the activity of digestive serine protease, and the verification of its functional activity can be helpful to further investigate the relationship between ApSerpin-FA72 and serine protease.
Service
E-mail this article
Add to my bookshelf
Add to citation manager
E-mail Alert
RSS
Articles by authors
ZHAO Yu
LIU Zhong-Yuan
Key wordsAnatolica polita borealis; serine protease inhibitor   prokaryotic expression   protein purification   functional verification     
Cite this article:   
ZHAO Yu,LIU Zhong-Yuan. Cloning, expression and functional verification of the serine protease inhibitor gene ApSerpin-FA72 from Anatolica polita borealis (Coleoptera: Tenebrionidae)[J]. ACTA ENTOMOLOGICA SINICA, 2017, 60(8): 857-864.
URL:  
http://www.insect.org.cn/EN/10.16380/j.kcxb.2017.08.002      or     http://www.insect.org.cn/EN/Y2017/V60/I8/857
 
No references of article
[1] LI Zhao-Qun, ZHANG Shuai, ZHOU Shu-Fen, LUO Jun-Yu, CUI Jin-Jie. Tissue expression profiling and ligandbinding properties of HarmOBP16 of the cotton bollworm, Helicoverpa armigera (Lepidoptera: Noctuidae)[J]. ACTA ENTOMOLOGICA SINICA, 2017, 60(8): 891-899.
[2] YANG Ji-Gui, PENG Li-Na, LIU Tai-Xing, WU Yun-Fei, DU Guo-Yu, ZHANG Qian, WANG Wei, PAN Min-Hui. Cloning and expression analysis of the Sox family gene BmDichaete in the silkworm, Bombyx mori[J]. ACTA ENTOMOLOGICA SINICA, 2017, 60(5): 513-522.
[3] ZHANG Yuan-Chen, Liu Xiang-Dong. Cloning and expression analysis of lysozyme gene AgLYS in the cotton-melon aphid,Aphis gossypii (Hemiptera: Aphididae)[J]. ACTA ENTOMOLOGICA SINICA, 2017, 60(2): 127-135.
[4] FANG Chi-Qin, ZHANG Xin-Xin, LIU Dan-Dan, LI Ke-Bin, ZHANG Shuai, CAO Ya-Zhong, FAN Dong, YIN Jiao. Cloning and functional analysis of an odorant-binding protein HparOBP15a gene from  Holotrichia parallela  (Coleoptera: Melolonthidae)[J]. ACTA ENTOMOLOGICA SINICA, 2016, 59(3): 260-268.
[5] WU Peng-Jie, JIN Hong-Yan, XU Jin, WU Jiang-Li, LI Yu-Shi, GUO Yue-Qin. Molecular cloning, sequence analysis and prokaryotic expression of SRP9 gene in the Chinese honeybee, Apis cerana cerana (Hymenoptera: Apidae)[J]. ACTA ENTOMOLOGICA SINICA, 2016, 59(12): 1332-1339.
[6] HAILIQIEMU AiLi, XU Xin, LIU Zhong-Yuan. Optimization of prokaryotic expression conditions and the antibacterial activity of a novel antimicrobial peptide from  Anatolica polita borealis (Coleoptera: Tenebrionidae)[J]. ACTA ENTOMOLOGICA SINICA, 2016, 59(1): 8-15.
[7] QIAN Kai, FENG Bo, WU Xu-Liang, LAO Cong, SHEN You-Lian, DU Yong-Jun. Cloning, sequence analysis, tissue expression profiling and prokaryotic expression of odorant binding protein genes MaltOBP2 and MaltOBP6 from Monochamus alternatus (Coleoptera: Cerambycidae)[J]. ACTA ENTOMOLOGICA SINICA, 2015, 58(5): 496-506.
[8] YIN Shu-Yan , YANG Chun-Hong , LIU Zhao-Yang , YU Qi , GAO Hui-Ju , CUI Wei-Zheng , LIU Qing-Xin . Preparation of the antigen protein of the atypical olfactory receptor Orco in the silkworm (Bombyx mori) with prokaryotic expression system[J]. ACTA ENTOMOLOGICA SINICA, 2015, 58(11): 1194-1200.
[9] JIA Xiao-Jian, HAO Shao-Dong, DU Yan-Li, ZHANG Min-Zhao, QIN Xiao-Chun, WANG Jin-Zhong, WANG Hai-Xiang, JI Wei-Rong. cDNA cloning, expression profiling and binding affinity assay of the pheromone binding protein Cpun-PBP1 in the yellow peach moth, Conogethes punctiferalis (Lepidoptera: Crambidae)[J]. ACTA ENTOMOLOGICA SINICA, 2015, 58(11): 1167-1176.
[10] LI Wen-Hai, HUANG Xing-Long, WANG Dun, FENG Ji-Nian. Molecular cloning and immunofluorescence localization of sex pheromone binding protein 2 from Atrijuglans hetaohei (Lepidoptera: Heliodinidae)[J]. ACTA ENTOMOLOGICA SINICA, 2015, 58(10): 1054-1062.
[11] ZHANG Zhi-Ke, WU Sheng-Yong, LEI Zhong-Ren. cDNA cloning, expression profiling and immunolocalization of a chemosensory protein in the western flower thrips,  Frankliniella occidentalis (Thysanoptera: Thtipidae)[J]. ACTA ENTOMOLOGICA SINICA, 2015, 58(1): 1-14.
[12] CHEN Wei, XU Wei-Hua. Molecular cloning, preparation of polyclonal antibody and expression detection of Wnt1 in the brain of diapausing and non-diapausing pupae of  Helicoverpa armigera (Lepidoptera: Noctuiadae)[J]. ACTA ENTOMOLOGICA SINICA, 2015, 58(1): 15-21.
[13] ZHOU Xiao-Qun, GAO Yan-Ling, ZHAO Kui-Jun, FAN Dong. cDNA cloning, sequence analysis and prokaryotic expression of a serine protease from the midgut of Heliothis viriplaca (Lepidoptera: Noctuidae)[J]. ACTA ENTOMOLOGICA SINICA, 2014, 57(9): 1008-1017.
[14] JI Ting, SHEN Fang, LIANG Qin, WU Li-Ming, LIU Zhen-Guo, LUO Yue-Xiong. Cloning, prokaryotic expression and tissue expression profiling of an OBP3 gene in the Chinese honeybee, Apis cerana cerana (Hymenoptera: Apidae)[J]. ACTA ENTOMOLOGICA SINICA, 2014, 57(8): 897-904.
[15] ZHAO Tong-Wei ZHANG Bing, Lü Wen-Jing, ZHAI Hui-Feng, WU Tao-Yan, ZHANG Shi-Yang, TANG Xiao-Wei, FENG Cong-Jing. Prokaryotic expression and characterization of prophenoloxidase from the Asian corn borer, Ostrinia furnacalis (Lepidoptera: Pyralidae)[J]. ACTA ENTOMOLOGICA SINICA, 2014, 57(7): 798-805.
Copyright © 2011 ACTA ENTOMOLOGICA SINICA
Supported by:Beijing Magtech