Cloning and expression analysis of lysozyme gene <em>AgLYS</em> in the cotton-melon aphid,<em>Aphis gossypii</em> (Hemiptera: Aphididae)
昆虫学报
       Home | About Journal | Editorial Board | Subscription | Instruction | Advertisement | Contact Us | Chinese
ACTA ENTOMOLOGICA SINICA  2017, Vol. 60 Issue (2): 127-135    DOI: doi:10.16380/j.kcxb.2017.02.001
RESEARCH PAPERS Current Issue | Next Issue | Archive | Adv Search  |  Next Articles  
Cloning and expression analysis of lysozyme gene AgLYS in the cotton-melon aphid,Aphis gossypii (Hemiptera: Aphididae)
ZHANG Yuan-Chen, LIU Xiang-Dong*
(College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China)
 Download: PDF (4184 KB)   HTML (1 KB)   Export: BibTeX | EndNote (RIS)      Supporting Info
Abstract 【Aim】 This study aims to clone the lysozyme gene of the cotton-melon aphid Aphis gossypii Glover, to construct the expression system of this gene in Escherichia coli, and to analyze its expression profiles in aphids of different day-old and reared on different host plants, so as to further research the biological function of this gene and to illustrate the relationship between lysozyme and symbionts in aphids. 【Methods】 The total RNA was extracted from apterous adults of A. gossypii and used as the template to clone lysozyme gene and to obtain the full-length cDNA sequence using RT-PCR and RACE techniques. The gene fragment of lysozyme gene in A. gossypii without signal peptide sequence was linked to the expression vector pET-30a and then transformed into E. coli to express. The qRT-PCR method was used to examine the relative expression levels of the lysozyme gene in A. gossypii of different day-old (1-19-day-old) and reared on different host plants (zucchini, cucumber and cowpea). 【Results】 A lysozyme gene was cloned from apterous aphids of A. gossypii and named AgLYS with the GenBank accession no. KY575341. The full-length cDNA of AgLYS is 680 bp, and its ORF is 516 bp which encodes 172 amino acid residues. A signal peptide sequence with 36 amino acid residues was found at the N-terminus. The phylogenetic analysis showed that the lysozyme of A. gossypii belongs to i-type and has 90% amino acid sequence identity with that of the pea aphid Acyrthosiphon pisum. AgLYS was expressed in E. coli, and the expressed protein has the molecular weight of approximate 20 kD. AgLYS could be expressed in 1-19-day-old A. gossypii aphids. Its expression levels were lower in 1-9-day-old aphids, but increased extremely significantly in 11-19-day-old aphids (P<0.01). The expression levels of AgLYS were extremely significantly higher in A. gossypii aphids reared on zucchini than in those reared on cucumber and cowpea (P<0.01). 【Conclusion】 Lysozyme i-type was found in A. gossypii, and AgLYS was cloned and expressed in E. coli. The expression of AgLYS is influenced by developmental stages of A.gossypii and host plants. It is inferred that AgLYS might be involved in the regulation of population density of symbionts in A.gossypii.
Service
E-mail this article
Add to my bookshelf
Add to citation manager
E-mail Alert
RSS
Articles by authors
ZHANG Yuan-Chen
Liu-Xiang-Dong
Key wordsAphis gossypii   lysozyme   gene cloning   gene expression   prokaryotic expression   host plant     
Cite this article:   
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.
URL:  
http://www.insect.org.cn/EN/doi:10.16380/j.kcxb.2017.02.001      or     http://www.insect.org.cn/EN/Y2017/V60/I2/127
 
No references of article
[1] QIN Hua-Wei, Men Xing-Yuan, Yu-Yi, Lu Zeng-Bin, Sun Ting-Lin, Zhou Xian-Hong, Li Li-Li. Effects of host plants on the cold hardiness of Athetis lepigone (Lepidoptera: Noctuidae) larvae[J]. ACTA ENTOMOLOGICA SINICA, 2017, 60(2): 205-210.
[2] CHEN Li-Peng, DU Yu-Ping, FANG Qi, YAO Hong-Wei, YE Gong-Yin. Molecular cloning, sequence analysis and expression profiling of vitellogenin receptor gene in the green rice leafhopper,  Nephotettix cincticeps  (Hemiptera: Cicadellidae)[J]. ACTA ENTOMOLOGICA SINICA, 2016, 59(9): 925-937.
[3] HUANG Ling, SUN Liang-Zhen, WANG Yong, RU Yu-Tao, Muhammad IRFAN, JIANG Yi-Ren, Shi-Sheng-Lin , YANG Rui-Sheng, LI Xi-Sheng, QIN Li. Changes in the expression of trehalose-6-phosphate synthase gene in Antheraea pernyi (Lepidoptera: Saturniidae) during pupal diapause termination[J]. ACTA ENTOMOLOGICA SINICA, 2016, 59(9): 938-947.
[4] QIN Jiang-Mei, CAI Li-Jun, ZHENG Li-Shuang, CHENG Xiao-Juan, YOU Min-Sheng. Identification and ligand binding characteristics of antennal binding protein PxylOBP31 in the diamondback moth, Plutella xylostella (Lepidoptera: Plutellidae)[J]. ACTA ENTOMOLOGICA SINICA, 2016, 59(8): 812-822.
[5] LIN Fang-Fei, ZUO Ya-Yun, ZHAO Qi, DUAN Xin-Le, LI Yu-Ting, CHEN Mao-Hua. Molecular cloning, characterization and induced expression analysis of ion channel gene RSC1 in Rhopalosiphum padi (Hemiptera: Aphididae)[J]. ACTA ENTOMOLOGICA SINICA, 2016, 59(7): 716-723.
[6] TIAN Yu, DU Juan, LI Shang-Wei, LI Jiao, WANG Shuang. Molecular cloning, characterization and expression analysis of trehalase genes in the rice leaf folder, Cnaphalocrocis medinalis (Lepidoptera: Pyralidae)[J]. ACTA ENTOMOLOGICA SINICA, 2016, 59(6): 602-612.
[7] LI Kai-Xia, MA En-Bo, ZHANG Jian-Zhen, ZHAO Yi-Dan, ZHANG Xu-Bo. Combination of Gal80ts and Gal4 flexibly manipulates the expression levels of UAS transgenes in Drosophila[J]. ACTA ENTOMOLOGICA SINICA, 2016, 59(5): 481-488.
[8] JIANG En-Shun, ZHU Yi, WANG Jiang-Yong. Biological effectiveness of extracts from Vitex negundo against Aphis gossypii (Homoptera: Aphididae)[J]. ACTA ENTOMOLOGICA SINICA, 2016, 59(5): 538-545.
[9] SI Feng-Ling, HE Zheng-Bo, CHEN Bin. Cloning and expression profiling of heat shock protein DaHSP23 gene in the winter and summer diapause
pupae of the onion maggot, Delia antiqua(Diptera: Anthomyiidae) (In English)
[J]. ACTA ENTOMOLOGICA SINICA, 2016, 59(4): 402-410.
[10] WANG Qian, BAO Wei-Fang, ZENG Juan, YANG Yi-Zhong, LU Yan-Hui. Tracking the trophic relationship between herbivorous insects and host plants by DNA-based technology[J]. ACTA ENTOMOLOGICA SINICA, 2016, 59(4): 472-480.
[11] CHENG Xiao-Juan, CAI Li-Jun, ZHENG Li-Shuang, QIN Jiang-Mei, HUANG Yu-Ping, YOU Min-Sheng . Cloning, expression profiling and binding characterization of the OBP2 gene in the diamondback moth, Plutella xylostella (Lepidoptera: Plutellidae)[J]. ACTA ENTOMOLOGICA SINICA, 2016, 59(4): 365-376.
[12] ZHANG Xiao-Hong, LI Yan-Li, ZHANG Kai-Jian, HOU Juan, YIN Hong. Cloning and expression analysis of the hexamerin subunit type 2 (Hex2) gene from the grasshopper Calliptamus italicus (Orthoptera: Catantopidae)[J]. ACTA ENTOMOLOGICA SINICA, 2016, 59(2): 156-163.
[13] LIU Yang, GAO He, LI Xue-Jiao, WU Jun-Xiang, CHENG Wei-Ning, ZHU-SALZMAN Ke-Yan. Changes in sorbitol content and the expression level of sorbitol dehydrogenase gene in Sitodiplosis mosellana (Diptera: Cecidomyiidae) larvae at different diapause stages[J]. ACTA ENTOMOLOGICA SINICA, 2016, 59(2): 119-126.
[14] 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.
[15] LIU Yi-Cong, WANG Ling, ZHANG You-Jun, XIE Wen, WU Qing-Jun, WANG Shao-Li. Abamectin resistance and expression of resistance-related genes in field populations of Tetranychus urticae (Acari: Tetranychidae) in China[J]. ACTA ENTOMOLOGICA SINICA, 2016, 59(11): 1199-1205.
Copyright © 2011 ACTA ENTOMOLOGICA SINICA
Supported by:Beijing Magtech