金小蜂是一类寄生性昆虫，靶向许多人类疾病传播的载体（如家蝇等），在害虫生物防治上具有重要地位。作为一个新的模式昆虫，近期公布的丽蝇蛹集金小蜂（Nasonia vitripennis）全基因组序列为研究寄生性昆虫天然免疫的分子和进化机制奠定了基础。

运用进化基因组学的方法，中科院动物研究所朱顺义研究员领导的团队从丽蝇蛹集金小蜂基因组中鉴定了44个抗微生物肽新基因，组建了*个寄生性昆虫抗微生物肽基因蓝图。进一步研究证实，金小蜂抗微生物肽基因在细菌攻击后转录本的表达水平显着上调。利用化学合成和遗传重组表达的蛋白，他们对其中不同类别的代表性序列进行了结构、功能和进化研究，确定了4个抗微生物肽基因的抗微生物活性，发现γ－core区域是防御肽Navidefensin2-2的抗菌活性表面。他们的结果还表明，基因重复和功能区域的正选择可能驱动了金小蜂防御肽基因家族的适应性进化。

此外，运用比较基因组学的方法，研究人员还发现，与同为膜翅目的意大利蜜蜂（Apis mellifera）相比，金小蜂直系同源的抗微生物肽基因发生了明显的变化。主要表现在基因数量扩张，蛋白质末端延长，功能域的串联重复和融合以及结构多样性改变等。他们发现，基因和外显子重复以及外显子改组是造成这类寄生性昆虫免疫防御分子复杂度增加的zui主要原因。

该系列研究工作的科学意义在于：1）在上建立了*个寄生性昆虫的全套抗微生物肽数据，为金小蜂天然免疫以及寄生和免疫的关系研究奠定了基础；2）该研究发展的快速鉴定抗微生物肽基因的计算基因组学策略，有望拓展到其它模式生物，包括人类抗微生物肽新基因的发现，这将加速人类对于抗微生物肽介导的天然免疫防御网络进化的研究。

推荐原文出处1：

Process Biochemistry doi:10.1016/j.procbio.2009.08.017

Characterization of a hymenoptaecin-like antimicrobial peptide in the parasitic wasp Nasonia vitripennis
Bin Gaoa and Shunyi Zhu, a,

a Group of Animal Innate Immunity, State Key Laboratory of Integrated Management of Pest Insects & Rodents, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China

Hymenoptaecin is a Hymenoptera insect-specific, glycine-rich antimicrobial peptide （AMP） found in non-parasitic bees. Here, we describe a unique hymenoptaecin-like gene （named nahymenoptaecin-1） in the parasitic wasp Nasonia vitripennis, which codes for a larger protein precursor with a carboxyl-terminal hymenoptaecin-like domain （HLD） similar to the bee hymenoptaecin. We recombinantly produced its full-length bioactive form as well as 1–33 and 34–98 fragments （named HLD-n and HLD-c, respectively）. Recombinant HLD exhibited activity against Gram-negative and Gram-positive bacteria at micromolar concentrations. Compared to the full-length peptide, HLD-c possessed similar potency in inhibiting the growth of Stenotrophomonus but had a narrower antibacterial spectrum, whereas HLD-n only displayed weak effect on Stenotrophomonus, suggesting that HLD-n is a crucial determinant for bacterial target selectivity while HLD-c represents its active unit for the whole molecule. Circular dichroism analysis combined with ab initio structure prediction by Robetta indicated that HLD-n adopts a random coil conformation whereas glycine-rich HLD-c forms a loose β-sheet structure. Relative to bee hymenoptaecin, the upstream region of HLD contains two accuray repeated proline-rich AMP-like peptides instead of an acidic propeptide. Such difference could be a consequence of exon shuffling of autonomous modules after speciation.

推荐原文出处2：

Dev Comp Immunol. PMID: 20097222

Identification and characterization of the parasitic wasp Nasonia defensins: positive selection targeting the functional region
Gao B, Zhu S.

Group of Animal Innate Immunity, State Key Laboratory of Integrated Management of Pest Insects & Rodents, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China.

Defensin is a crucial component of innate immunity highly conserved across different insect orders. Here, we report identification and characterization of defensins in the parasitic wasp Nasonia （Hymenoptera: Pteromalidae）. In comparison with those in the non-parasitic insect Apis mellifera, two different subtypes of defensins （defensin1 and defensin2） have undergone independent gene duplication to create a mutigene family of five members （named 1-1, 1-2, 2-1, 2-2 and 2-3） in the Nasonia lineage. Such duplication occurred before the divergence of three sibling species （N. vitripennis, N. giraulti and N. longicornis） and the duplicated genes was subsequently subjected to positive selection at the amino-terminal loop and the gamma-core region. RT-PCR identified that only the subtype 1 of defensins were constitutively expressed in the N. vitripennis adult stage and none of the five defensins was expressed in other developmental stages （i.e. the infected Musca domestica pupae）. A functional form of 2-2 in N. vitripennis （named navidefensin2-2） was produced in Escherichia coli by an on-column refolding approach. The recombinant peptide presented a typical defensin structure, as identified by CD analysis, and selectively inhibited the growth of two Gram（+） bacteria at low micromolar concentrations. The bioactive surface responsible for antibacterial activity of navidefensin2-2 was identified in the gamma-core region of this molecule. Positive selection targeting the antibacterial region of defensins could be a consequence of evolutionary arms race between Nasonia and its pathogens. Copyright 2010 Elsevier Ltd. All rights reserved.

上海劲马生物（）推荐原文出处3：

BMC Genomics 2010, 11:187doi:10.1186/1471-2164-11-187

Antimicrobial peptide-like genes in Nasonia vitripennis: a genomic perspective
Caihuan Tian1 , Bin Gao1 , Qi Fang2 , Gongyin Ye2  and Shunyi Zhu1

1 Group of Animal Innate Immunity, State Key Laboratory of Integrated Management of Pest Insects & Rodents, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, PR China
2 State Key Laboratory of Rice Biology, Institute of Insect Sciences, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou 310029, PR China

Background
Antimicrobial peptides （AMPs） are an essential component of innate immunity which can rapidly respond to diverse microbial pathogens. Insects, as a rich source of AMPs, attract great attention of scientists in both understanding of the basic biology of the immune system and searching molecular templates for anti-infective drug design. Despite a large number of AMPs have been identified from different insect species, little information in terms of these peptides is available from parasitic insects.

Results
By using integrated computational approaches to systemically mining the Hymenopteran parasitic wasp Nasonia vitripennis genome, we establish the first AMP repertoire whose members exhibit extensive sequence and structural diversity and can be distinguished into multiple molecular types, including insect and fungal defensin-like peptides （DLPs） with the cysteine-stabilized α-helical and β-sheet （CSαβ） fold; Pro- or Gly-rich abaecins and hymenoptaecins; horseshoe crab tachystatin-type AMPs with the inhibitor cystine knot （ICK） fold; and a linear α-helical peptide. Inducible expression pattern of seven N. vitripennis AMP genes were verified, and two representative peptides were synthesized and functionally identified to be antibacterial. In comparison with Apis mellifera （Hymenoptera） and several non-Hymenopteran model insects, N. vitripennis has evolved a complex antimicrobial immune system with more genes and larger protein precursors. Three classical strategies that are likely responsible for the complexity increase have been recognized: 1） Gene duplication; 2） Exon duplication; and 3） Exon-shuffling.

Conclusion
The present study established the N. vitripennis peptidome associated with antimicrobial immunity by using a combined computational and experimental strategy. As the first AMP repertoire of a parasitic wasp, our results offer a basic platform for further studying the immunological and evolutionary significances of these newly discovered AMP-like genes in this class of insects.