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[1]张洁,王国栋.杂色鲍C型溶菌酶基因的克隆及表达分析[J].集美大学学报(自然科学版),2020,25(5):328-335.
　ZHANG Jie,WANG Guodong.Molecular Cloning and Expression Analysis of C-Type Lysozyme Gene in Haliotis diversicolor[J].Journal of Jimei University,2020,25(5):328-335.
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杂色鲍C型溶菌酶基因的克隆及表达分析(PDF)
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《集美大学学报（自然科学版）》[ISSN:1007-7405/CN:35-1186/N]

卷:
第25卷
期数:
2020年第5期
页码:
328-335
栏目:
水产、食品与生物工程
出版日期:
2020-09-30

文章信息/Info

Title:
Molecular Cloning and Expression Analysis of C-Type Lysozyme Gene in Haliotis diversicolor
作者:
张洁1; 2; 王国栋1; 2
(1.集美大学水产学院,福建 厦门 361021 ;2.农村农业部北方海水养殖重点实验室,福建 厦门 361021 )
Author(s):
ZHANG Jie1; 2; WANG Guodong1; 2
(1.Fisheries College,Jimei University,Xiamen 361021,China;2.Key Laboratory of Mariculture for North of Agriculture,Xiamen 361021,China)
关键词:
杂色鲍; C型溶菌酶; 基因克隆; 表达分析
Keywords:
Haliotis diversicolor ; C-type lysozyme; gene clone; expression analysis
分类号:
-
DOI:
-
文献标志码:
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摘要:
为了了解C型溶菌酶与杂色鲍（Haliotis diversicolor）的抗菌效应关系，通过SMART-RACE技术克隆到一条杂色鲍C型溶菌酶基因（HdLysC）全长cDNA，经生物信息学分析及应激表达分析得到以下结果：1）基因全长591 bp，开放阅读框441 bp，编码146个氨基酸，有1个LYZ1功能域（20—146aa)、1个保守基序（G/T）SSDYGLFQINS和两个催化位点(Glu52和Asp70)。2）多重比对和系统发育分析表明,HdLysC与亲缘关系较近的盘鲍（Haliotis discus discus）C2型溶菌酶基因的同源性最高。3)组织表达差异实验表明，HdLysC基因在各组织中均有表达且差异显著（P<0.01），在外套膜组织中的表达量最高，而在血淋巴中的表达量最低。4）实时荧光定量PCR（qRT-PCR）实验表明，HdLysC能够响应副溶血弧菌感染，其表达水平随着感染的不同时相而变动。在副溶血弧菌感染4 h后，HdLysC在血淋巴组织中表达量增加（未达到显著差异），而在鳃组织中表达量有所下降；感染12 h后，HdLysC在各组织中的表达量都有明显的增加，呈极显著差异（P<0.01），尤其在血淋巴中增加最多；感染24 h后，其表达量在各组织中都有所降低。
Abstract:
To understand the relationship between C-type lysozyme in the Haliotis diversicolor and the antibacterial effect of heterosorbide，the full-length of C-type lysozyme gene cDNA sequence was cloned by SMART RACE in H.diversicolor.The results showed that the full-length cDNA sequence is 591 bp，with a 441 bp open reading frame.The deduced protein contained 146 amino acids，with a LYZ1 domain (20-146aa)，a conserved motif (G/T) SSDYGLFQINS and two catalytic sites (Glu52 and Asp 70).Multiple alignments and phylogenetic analysis showed that the C-type lysozyme gene of H.diversicolor had the highest homology the C-2 type lysozyme gene of H.discus discus.The experimental results showed the C-type lysozyme gene in H.diversicolor was expressed in all detected tissues.The highest expression of C-type lysozyme gene was detected in the mantle (P<0.01)，and the lowest expression in hemolymph.Real-time quantitative PCR results showed that the expression of C-type lysozyme gene in H.diversicolor increased in blood lymphoid，the expression in gill was declined at 4 h post challenged by Vibrio parahaemolyticus.The expression of C-type lysozyme gene in different tissues increased significantly (P<0.01) at 12 h post challenged，especially in the hemolymph.The expression decreased in various tissues at 24 h post challenged.These results showed C-type lysozyme gene in H.diversicolor could respond to the infection of V.parahaemolyticus，while the expression fluctuated in different time after infection.

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更新日期/Last Update: 2020-11-04