[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()
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《集美大学学报(自然版)》[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、食品与生物技术
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12王国栋12
(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 Jie12WANG Guodong12
(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 lysozymegene cloneexpression analysis
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.
更新日期/Last Update: 2020-11-04