Cloning，Expression and Sequence Analysis of Musca domestica Antifungal Peptide-1 and Musca domestica Lysozyme

doi:10.13560/j.cnki.biotech.bull.1985.2015.05.031

Abstract

Abstract: The aim of this study is to have bioinformatics analysis of Musca domesitca antifungal peptide-1（MAF-1）and lysozymeb（LMZ）, also clone fused MAF-1-LMZ gene and have expression analysis of it. The encoding sequences of MAF-1 and LMZ were fetched from GenBank, and the structures and functions of 2 proteins were analyzed and predicted. The gene MAF-1-LMZ was amplified by polymerase chain reaction（PCR）, then was ligated into pET 28a and transformed into E. coli Origmi（DE3）competent cell, and induced with IPTG. The fusion protein MAF-1-LMZ in the expression vector was analyzed by SDS-PAGE. The activity of the protein was validated by methods of small test tube and doubling dilution. The open reading frame of the MAF-1-LMZ was 969 bp, encoding a putative protein consisting of 322 amino acids with a predicted molecular weight of 35 468.6 Da and pI of 8.31, indicating the gene expressed in E. coli Origmi（DE3）successfully. The purified target protein had the antifungal activity.

Key words: Musca domestica, antifungal peptide-1 with Lysozyme, recombinant expression, sequence analysis

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Peng Chuanlin , Wei Chuanchuan, Wu Jianwei, Wang Yu, Xiu Jiangfan, Shang Xiaoli, Zhao Xuejun. Cloning，Expression and Sequence Analysis of Musca domestica Antifungal Peptide-1 and Musca domestica Lysozyme[J]. Biotechnology Bulletin, 2015, 31(5): 200-205.

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Cloning，Expression and Sequence Analysis of Musca domestica Antifungal Peptide-1 and Musca domestica Lysozyme

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[1]	RAO Zi-huan, XIE Zhi-xiong. Isolation and Identification of a Cellulose-degrading Strain of Olivibacter jilunii and Analysis of Its Degradability [J]. Biotechnology Bulletin, 2023, 39(8): 283-290.
[2]	PAN Guo-qiang, WU Si-yuan, LIU Lu, GUO Hui-ming, CHENG Hong-mei, SU Xiao-feng. Construction and Preliminary Analysis of Verticillim dahliae Mutant Library [J]. Biotechnology Bulletin, 2023, 39(5): 112-119.
[3]	YOU Chui-huai, XIE Jin-jin, ZHANG Ting, CUI Tian-zhen, SUN Xin-lu, ZANG Shou-jian, WU Yi-ning, SUN Meng-yao, QUE You-xiong, SU Ya-chun. Identification of the Lipoxygenase Gene GeLOX1 and Expression Analysis Under Low Temperature Stress in Gelsmium elegans [J]. Biotechnology Bulletin, 2023, 39(11): 318-327.
[4]	CHANG Qing, SHU Yue-rong, WANG Wen-tao, JIANG Hao, YAN Quan-de, QIAN Zheng, GAO Xue-chun, WU Jin-hong, ZHANG Yong. Heterologous Expression and Characterization of Endo-type Alginate Lyase from Yeosuana marina sp. JLT21 [J]. Biotechnology Bulletin, 2022, 38(2): 123-131.
[5]	CAO Ru-fei, LI Ze-xuan, XU Huan, ZHANG Sha, ZHANG Min-min, DAI Feng, DUAN Xiao-lei. Expression,Purification,and Crystallization of Pif1 Helicase from Bacteroides fragilis [J]. Biotechnology Bulletin, 2021, 37(9): 180-190.
[6]	DUAN Xu-guo, ZHANG Yu-hua, HUANG Ting-ting, DING Qian, LUAN Shu-yue, ZHU Qiu-yu. Synergetic Enhancing the Soluble Expression of Thermotoga maritima α-Glucan Phosphorylase by Chemical Chaperones and Induction Condition Optimization [J]. Biotechnology Bulletin, 2021, 37(8): 233-242.
[7]	LIN Yan-li, QIN Jian-bing, WU Xiang, WANG Yan-yan, PAN You-zhao, LIU Zhong-yu. Cloning and Activity Analysis of PcMYB1 Promoter from Polygonum cuspidatum [J]. Biotechnology Bulletin, 2021, 37(5): 48-55.
[8]	YAO Qiong, QUAN Lin-fa, XU Shu, DONG Yi-zhi, LI Wen-jing, CHI Yan-yan, CHEN Bing-xu. Gene Cloning of 2 Octopamine Receptors from Eocanthecona furcellata and Effects of Chemical Pesticide on Its Expression [J]. Biotechnology Bulletin, 2021, 37(10): 152-152.
[9]	ZHAO Zhen, WANG Sha-sha, LÜ Xing-xing, TAO Yan, XIE Jing, QIAN Yun-fang. Heterologous Expression of Cyclina sinensis Mytimacin Antibacterial Peptide Based on Recombinant Pichia pastoris [J]. Biotechnology Bulletin, 2020, 36(5): 150-158.
[10]	GAO Yun-shan, LIU Dan-dan, XU Jun-lin, SANG Yu-nong, LIANG Xia-xia, LIU Jian-xin, WANG Wen-bin. Recombinant Expression and Immunogenicity Analysis of the Porin Protein OmpF of Aeromonas hydrophila [J]. Biotechnology Bulletin, 2019, 35(9): 234-243.
[11]	ZHANG Ya-li, TAO Yan, XIE Jing, QIAN Yun-fang. Recombinant Expression of Mytilus coruscus Mytilin-1 Mature Peptide in Pichia pastoris and Its Antibacterial Activity [J]. Biotechnology Bulletin, 2019, 35(7): 54-60.
[12]	LIU Huan, XU Ling-ling, LI Jiang. Identification and Carbon Metabolic Analysis of a Mold Isolated from Uranium Acid Leaching Solution [J]. Biotechnology Bulletin, 2018, 34(4): 208-213.
[13]	YANG Yuan-xiao, ZI Xiang-dong. Cloning of KISS-1 Gene in Yak and Its Expression in Reproductive Axis [J]. Biotechnology Bulletin, 2018, 34(3): 142-149.
[14]	JIN Qiao, LIU Xia, GAN Zhi-Kai. Sequencing and Analyzing Mitochondrial Genome of Cacopsylla coccinea [J]. Biotechnology Bulletin, 2018, 34(11): 127-135.
[15]	GUO Ya-fei, WANG Jun-ya, GUO Fei, NI De-jiang. Cloning and Expression Analysis of CsDXS1 Gene Encoding 1-Deoxy-D-Xylulose-5-Phosphate Synthase in Camellia sinensis [J]. Biotechnology Bulletin, 2018, 34(1): 144-152.