Journal of Guangxi Normal University(Natural Science Edition) ›› 2021, Vol. 39 ›› Issue (1): 156-164.doi: 10.16088/j.issn.1001-6600.2020083102

Previous Articles    

Cloning and Bioinformatics Analysis of LAP Gene from Pseudomonas

GUO Chen1, ZHOU Fei 2,3, HAN Biao1, PAN Cui1, WU Jiemin1, YANG Ting2,3, SHANG Changhua2,3*   

  1. 1. Scientific Research Academy of Guangxi Environmental Protection, Nanning Guangxi 530022, China;
    2. College of Life Science, Guangxi Normal University, Guilin Guangxi 541006, China;
    3. Key Laboratory of Ecology of Rare and Endangered Species and Environmental Protection (Guangxi Normal University), Ministry of Education, Guilin Guangxi 541006, China
  • Received:2020-08-31 Revised:2020-09-22 Online:2021-01-25 Published:2021-03-27

PDF (PC)

31

Abstract: To explore the molecular structure and functional characteristics of LAP gene encoding Leucine aminopeptidases (LAPs) and its role in the process of growth and development, the Pseudomonas Cr13 LAP gene was cloned and characterized by bioinformatics in this study. Firstly, the core fragment of LAP was cloned. Secondly, the full-length DNA of LAP gene was cloned by two Genome Walking. Lastly, LAP gene was analyzed by bioinformatics software. The result showed that the full-length DNA of LAP gene was 1 491 bp in Pseudomonas Cr13. Bioinformatics analysis indicated that the LAP gene was located in the mitochondrion. The LAP gene had a conserved cytoplasmic aminopeptidase motif NTDAEGRL. The Pseudomonas Cr13 LAP promoter (426 bp) contained the basic elements such as TATA-Box and CAAT-box and some potential cis-acting element. Cloning of full-length DNA laid a good foundation for further research on LAP gene function. Cloning and characterization of the promoter region of Pseudomonas sp. Cr13 LAP gene provided substantial basis for further research on the mechanism of regulation and expression of LAP gene.

Key words: Pseudomonas, leucine aminopeptidase, genome walking, bioinformatics

CLC Number: 

  • Q933
[1] CHAO W S, GU Y Q, PAUTOT V V, et al. Leucine aminopeptidase RNAs, proteins, and activities increase in response to water deficit, salinity, and the wound signals systemin, methyl jasmonate, and abscisic acid[J]. Plant Physiology, 1999, 120(4): 979-992. DOI:10.1104/pp.120.4.979.
[2] MURPHY A, PEER W A, TAIZ L. Regulation of auxin transport by aminopeptidases and endogenous flavonoids[J]. Planta, 2000, 211(3): 315-324. DOI:10.1007/s004250000300.
[3] ISHIZAKI T, TOSAKA A, NARA T, et al. Leucine aminopeptidase during meiotic development[J]. European Journal of Biochemistry, 2002, 269(3):826-832. DOI:10.1046/j.0014-2956.2001.02713.x.
[4] MATSUI M, FOWLER J H, WALLING L L. Leucine aminopeptidases: diversity in structure and function[J]. Biological Chemistry, 2006, 387(12):1535-1544. DOI:10.1515/BC.2006.191.
[5] ZHANG S, CAI X, LUO X, et al. Molecular cloning and characterization of leucine aminopeptidase gene from Taenia pisiformis[J]. Experimental Parasitology, 2018, 186:1-9. DOI:10.1016/j.exppara.2018.01.012.
[6] WU M, YAN M, XU J, et al. Expression, tissue localization and serodiagnostic potential of Echinococcus granulosus leucine aminopeptidase[J]. International Journal of Molecular Sciences, 2018, 19(4):1063. DOI:10.3390/ijms19041063.
[7] HUANG S, WU Y, ZENG F, et al. A turn-on fluorescence probe based on aggregation-induced emission for leucine aminopeptidase in living cells and tumor tissue[J]. Analytica Chimica Acta, 2018, 1031:169-177. DOI:10.1016/j.aca.2018.05.032.
[8] JIA H, NISHIKAWA Y, LUO Y, et al. Characterization of a leucine aminopeptidase from Toxoplasma gondii[J]. Molecular and Biochemical Parasitology, 2010, 170(1):1-6. DOI:10.1016/j.molbiopara. 2009.11.005.
[9] XI H, TIAN Y, ZHOU N, et al. Characterization of an N-glycosylated Bacillus subtilis leucine aminopeptidase expressed in Pichia pastoris[J]. Journal of Basic Microbiology, 2015, 55(2):236-246. DOI:10.1002/jobm.201400368.
[10] SINGH A K, SINGH R, TOMAR D, et al. The leucine aminopeptidase of Staphylococcus aureus is secreted and contributes to biofilm formation[J]. International Journal of Infectious Diseases, 2012, 16(5): e375-e381. DOI:10.1016/j.ijid.2012.01.009.
[11] JAROCKI V M, SANTOS J, TACCHI J L, et al. MHJ_0461 is a multifunctional leucine aminopeptidase on the surface of Mycoplasma hyopneumoniae[J]. Open Biology, 2015, 5(1):140175. DOI:10.1098/rsob.140175.
[12] 向照举, 李小璐, 李治.假单胞菌属合成环脂肽分子的研究进展[J].中国生物制品学杂志, 2014,27(7):974-980. DOI:10.13200/j.cnki.cjb.000356.
[13] 杨光富, 魏云林. 假单胞菌研究现状及应用前景[J]. 生物技术通报, 2011(1):37-39,49. DOI:10.13560/j.cnki.biotech.bull.1985.2011.01.005.
[14] 查代明, 张炳火, 李汉全, 等. 假单胞菌属脂肪酶的分子生物学研究进展[J]. 中国生物工程杂志, 2015, 35(9):114-121. DOI:10.13523/j.cb.20150916.
[15] MOORE B D. Bifunctional and moonlighting enzymes: lighting the way to regulatory , control[J]. Trends in Plant Science, 2004, 9(5):221-228. DOI:10.1016/j.tplants.2004.03.005.
[16] RAWLINGS N D, BARRETT A J, BATEMAN A. MEROPS: the peptidase database[J]. Nucleic Acids Research, 2010, 38:D227-D233. DOI:10.1093/nar/gkp971.
[17] 胡健健, 陈玉磊, 段雪昆, 等. 皱纹盘鲍亮氨酸氨肽酶基因cDNA克隆及表达分析[J]. 集美大学学报(自然科学版), 2017, 22(5):31-39. DOI:10.19715/j.jmuzr.2017.05.005.
[18] 刘登冉, 赵丽, 陈月异, 等. 巴西橡胶树胶乳亮氨酸氨肽酶基因克隆及表达[J]. 热带作物学报, 2008, 29(4):403-409. DOI:10.3969/j.issn.1000-2561.2008.04.001.
[19] 阮文斌, 董迎辉, 高晓艳, 等. 文蛤亮氨酸氨肽酶LAP3的基因克隆及其在幼体不同发育时期、成体不同组织的表达分析[J]. 海洋学报, 2017, 39(2):96-104. DOI:10.3969/j.issn.0253-4193.2017.02.009.
[20] SHAPTER F M, WATERS D L. Genome walking[J]. Methods in Molecular Biology, 2014, 1099:133-146. DOI:10.1007/978-1-62703-715-0_12.
[21] LEONI C, VOLPICELLA M, DE LEO F, et al. Genome walking in eukaryotes[J]. The FEBS Journal, 2011, 278(21): 3953-3977. DOI:10.1111/j.1742-4658.2011.08307.x.
[22] 韩琪, 胡波, 李升云, 等. 斜纹夜蛾气味结合蛋白基因GOBP2转录调控区的克隆及活性分析[J]. 南京农业大学学报, 2019, 42(6):1059-1066. DOI:10.7685/jnau.201904004.
[23] 郭丽娜, 赵慧婷, 任有蛇, 等. 中华蜜蜂气味受体AcerOr1基因启动子克隆及序列分析[J]. 山西农业科学, 2020, 48(4):623-626. DOI:10.3969/j.issn.1002-2481.2020.04.32.
[24] 周惠强, 陈凯棱, 舒琥, 等. 光倒刺鲃mc5r基因的克隆及对饥饿的响应[J]. 南方水产科学, 2019, 15(4):99-106. DOI:10.12131/20190027.
[25] 梁明丽, 刘波, 张伟. 巴斯德毕赤酵母鼠李糖代谢基因LRA3功能及其启动子顺式作用元件的研究[J]. 生物技术进展, 2019, 9(5):518-526. DOI:10.19586/j.2095-2341.2019.0031.
[26] 冯靖, 杨灿, 卢娟芳, 等. 杏CCD1和CCD4启动子克隆及顺式作用元件分析[J]. 园艺学报, 2020, 47(5):939-952. DOI:10.16420/j.issn.0513-353x.2019-0610.
[1] JIANG Shaofeng , HUANG Jinqing , YU Xiaoyu , LI Yunfei , LU Zujun. Construction and Characterization of the GenomicLibrary of Klebsiella oxytoca [J]. Journal of Guangxi Normal University(Natural Science Edition), 2016, 34(2): 151-157.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
[1] YANG Liting, LIU Xuecong, FAN Penglai, ZHOU Qihai. Research Progress in Vocal Communication of Nonhuman Primates in China[J]. Journal of Guangxi Normal University(Natural Science Edition), 2021, 39(1): 1 -9 .
[2] BIN Shiyu, LIAO Fang, DU Xuesong, XU Yilan, WANG Xin, WU Xia, LIN Yong. Research Progress on Cold Tolerance of Tilapia[J]. Journal of Guangxi Normal University(Natural Science Edition), 2021, 39(1): 10 -16 .
[3] LIU Jing, BIAN Xun. Characteristics of the Orthoptera Mitogenome and Its Application[J]. Journal of Guangxi Normal University(Natural Science Edition), 2021, 39(1): 17 -28 .
[4] LI Xingkang, ZHONG Enzhu, CUI Chunyan, ZHOU Jia, LI Xiaoping, GUAN Zhenhua. Monitoring Singing Behavior of Western Black Crested Gibbon (Nomascus concolor furvogaster)[J]. Journal of Guangxi Normal University(Natural Science Edition), 2021, 39(1): 29 -37 .
[5] HE Xinming, XIA Wancai, BA Sang, LONG Xiaobin, LAI Jiandong, YANG Chan, WANG Fan, LI Dayong. Grooming Strategies of Resident Males with Different Number of Mates in Yunnan Snub-nosed Monkeys (Rhinopithecus bieti)[J]. Journal of Guangxi Normal University(Natural Science Edition), 2021, 39(1): 38 -44 .
[6] FU Wen, REN Baoping, LIN Jianzhong, LUAN Ke, WANG Pengcheng, WANG Bing, LI Dayong, ZHOU Qihai. Jiyuan Taihang Mountain Macaque Population and Conservation Status[J]. Journal of Guangxi Normal University(Natural Science Edition), 2021, 39(1): 45 -52 .
[7] ZHENG Jingjin, LIANG Jipeng, ZHANG Kechu, HUANG Aimian, LU Qian, LI Youbang, HUANG Zhonghao. White-headed Langurs Select Foods Based on Woody Plants' Dominances[J]. Journal of Guangxi Normal University(Natural Science Edition), 2021, 39(1): 53 -64 .
[8] YANG Chan, WAN Yaqiong, HUANG Xiaofu, YUAN Xudong, ZHOU Hongyan, FANG Haocun, LI Dayong, LI Jiaqi. Activity Rhythm of Muntiacus reevesi Based on Infrared Camera Technology[J]. Journal of Guangxi Normal University(Natural Science Edition), 2021, 39(1): 65 -70 .
[9] LI Youbang, NONG Juanli, YANG Wanlin, ZHAO Jiajia, ZHU Qiqi. Daily Activity Patterns of Two Sympatric Squirrels Callosciurus erythraeus and Dremomys rufigenis in Nonggang, Guangxi, China[J]. Journal of Guangxi Normal University(Natural Science Edition), 2021, 39(1): 71 -78 .
[10] CHENG Rui, HE Mingxian, ZHONG Chunying, LUO Shuyi, WU Zhengjun. Comparison of Swimming Ability between Wild and Captive Breeding Shinisaurus crocodilurus[J]. Journal of Guangxi Normal University(Natural Science Edition), 2021, 39(1): 79 -86 .
Copyright © Journal of Guangxi Normal University(Natural Science Edition), All Rights Reserved.
Tel: 0773-5857325　E-mail: gxsdzkb@mailbox.gxnu.edu.cn
Powered by Beijing Magtech Co. Ltd