水生生物学报
水生生物學報
수생생물학보
ACTA HYDROBIOLOGICA SINICA
2014年
3期
563-570
,共8页
孙敬敬%刘慧慧%周世权%王信超%范美华%申望%廖智
孫敬敬%劉慧慧%週世權%王信超%範美華%申望%廖智
손경경%류혜혜%주세권%왕신초%범미화%신망%료지
厚壳贻贝%抗菌肽%几丁质结合结构域%荧光定量PCR
厚殼貽貝%抗菌肽%幾丁質結閤結構域%熒光定量PCR
후각이패%항균태%궤정질결합결구역%형광정량PCR
Mytilus coruscus%Antibacterial peptide%Chitin-biding domain%RT-PCR
厚壳贻贝(Mytilus coruscus)广泛分布于我国东部海域,其体内富含各种抗菌肽分子,是研究软体动物免疫防御机制以及开发抗菌肽来源的新型生物抗生素的重要对象。采用多步反相高效液相色谱对厚壳贻贝血清进行分离纯化,获得一种分子量为6261.55 D的具有抗菌活性的多肽成分;经多肽N端测序和基因克隆,结果表明该抗菌肽由55个氨基酸残基构成,含6个半胱氨酸并形成三对二硫键。结构域分析表明该抗菌肽具有几丁质结合结构域(Chitin-biding domain),因此将该抗菌肽命名为mytichitin-A。Mytichitin-A对革兰氏阳性菌具有较强的抑制作用,同时对真菌及革兰氏阴性菌也具有抑制作用。荧光定量 PCR 检测表明, mytichitin-A主要在厚壳贻贝的性腺组织中表达且在细菌诱导后12h其表达量达到峰值。研究为深入了解厚壳贻贝抗菌肽的分子多样性及免疫机制奠定了基础。
厚殼貽貝(Mytilus coruscus)廣汎分佈于我國東部海域,其體內富含各種抗菌肽分子,是研究軟體動物免疫防禦機製以及開髮抗菌肽來源的新型生物抗生素的重要對象。採用多步反相高效液相色譜對厚殼貽貝血清進行分離純化,穫得一種分子量為6261.55 D的具有抗菌活性的多肽成分;經多肽N耑測序和基因剋隆,結果錶明該抗菌肽由55箇氨基痠殘基構成,含6箇半胱氨痠併形成三對二硫鍵。結構域分析錶明該抗菌肽具有幾丁質結閤結構域(Chitin-biding domain),因此將該抗菌肽命名為mytichitin-A。Mytichitin-A對革蘭氏暘性菌具有較彊的抑製作用,同時對真菌及革蘭氏陰性菌也具有抑製作用。熒光定量 PCR 檢測錶明, mytichitin-A主要在厚殼貽貝的性腺組織中錶達且在細菌誘導後12h其錶達量達到峰值。研究為深入瞭解厚殼貽貝抗菌肽的分子多樣性及免疫機製奠定瞭基礎。
후각이패(Mytilus coruscus)엄범분포우아국동부해역,기체내부함각충항균태분자,시연구연체동물면역방어궤제이급개발항균태래원적신형생물항생소적중요대상。채용다보반상고효액상색보대후각이패혈청진행분리순화,획득일충분자량위6261.55 D적구유항균활성적다태성분;경다태N단측서화기인극륭,결과표명해항균태유55개안기산잔기구성,함6개반광안산병형성삼대이류건。결구역분석표명해항균태구유궤정질결합결구역(Chitin-biding domain),인차장해항균태명명위mytichitin-A。Mytichitin-A대혁란씨양성균구유교강적억제작용,동시대진균급혁란씨음성균야구유억제작용。형광정량 PCR 검측표명, mytichitin-A주요재후각이패적성선조직중표체차재세균유도후12h기표체량체도봉치。연구위심입료해후각이패항균태적분자다양성급면역궤제전정료기출。
The researches on antibacterial peptides from Mytilus coruscus, an important mytilus on aquiculture, have significant value that helping people to understand the mechanism of innate immune system of this mussel. By using multi-dimensional reverse phase high performance liquid chromatography (RP-HPLC), a novel antimicrobial peptide was isolated from Mytilus coruscus serum. The mass and the N-terminal sequence of this peptide were analyzed by a combination of Mass Spectrometry and Edman degradation. This peptide consisted 55 amino acids and had a 6621.55 D with 6 Cysteines presented in its sequence forming 3 disulfide bonds. A chitin-biding domain was detected by domain analysis and the peptide was named mytichitin-A therefore. The mRNA transcripts of mytichitin-A were mainly detected in gonad, which indicated that mytichitin-A were specifically synthesized and stored in genital system. The expression level of mytichitin-A in gonad was up-regulated and reached to the highest level at 12h after bacterial challenge, which was 9-fold increase compared to that of the control group. These results indicated that mytichitin-A was involved in the host immune response against bacterial infection and might contribute to the clearance of invading bacteria.