Chiou Tzu-Ting, Lu Jenn-Kan, Wu Jen-Leih, Chen Thomas T, Ko Chi-Fong, Chen Jiann-Chu
Department of Aquaculture, College of Life Sciences, National Taiwan Ocean University, Keelung, 202, Taiwan, ROC.
Dev Comp Immunol. 2007;31(2):132-42. doi: 10.1016/j.dci.2006.05.007. Epub 2006 Jun 15.
A penaeidin family, mo-penaeidin was cloned from the haemocytes of tiger shrimp Penaeus monodon using genomic polymerase chain reaction (PCR) by gene specific primers. Analysis of nucleotide sequence revealed that this mo-penaeidin consists of 1348 bp containing one intron (680 bp) and two exons (210 and 458 bp). It has an open reading frame (ORF) of 222 p, which encodes a protein of 74 amino acids including a signal peptide of 19 amino acids. The calculated molecular mass of the mature protein (55 amino acids) is 6.059 kDa with an estimated pI of 9.3. The deduced amino acid sequence of mo-penaeidin has similarity to that of penaeidin from Fenneropenaeus chinensis (73%), Farfantepenaeus paulensis (66%), Litopenaeus schmitti (53-67%), L. stylirostris (50-67%), L. setiferus (50-62%), L. vannamei (44-66%), and Marsupenaeus japonicus (33%), respectively. Phylogenetic tree analysis indicated that penaeidin (including mo-penaeidin, penaeidin, and penaeidin 5, 2, 3k, 3c1) of P. monodon is distinct from penaeidin 1, penaeidin 2, penaeidin 3 and penaeidin 4 of other penaeid shrimps. The mo-penaeidin mRNA was detected in various tissues including ovary and mandibular organ. The mo-penaeidin mRNA was present in one cell to postlarva stage with higher level at nauplius I (9h post hatching) and higher expression during the intermoult stage indicating an early innate immunity and different immunity at moulting stage.
使用基因特异性引物通过基因组聚合酶链反应(PCR)从斑节对虾的血细胞中克隆出一个对虾抗菌肽家族成员——斑节对虾对虾抗菌肽(mo-penaeidin)。核苷酸序列分析表明,该mo-penaeidin由1348个碱基对组成,包含一个内含子(680个碱基对)和两个外显子(210和458个碱基对)。它有一个222个碱基对的开放阅读框(ORF),编码一个74个氨基酸的蛋白质,其中包括一个19个氨基酸的信号肽。成熟蛋白(55个氨基酸)的计算分子量为6.059 kDa,估计的pI为9.3。推导的mo-penaeidin氨基酸序列与中国明对虾的对虾抗菌肽(73%)、巴西对虾(66%)、史氏对虾(53 - 67%)、斯氏对虾(50 - 67%)、长缝对虾(50 - 62%)、凡纳滨对虾(44 - 66%)和日本囊对虾(33%)的对虾抗菌肽序列分别具有相似性。系统发育树分析表明,斑节对虾的对虾抗菌肽(包括mo-penaeidin、对虾抗菌肽以及对虾抗菌肽5、2、3k、3c1)与其他对虾科对虾的对虾抗菌肽1、对虾抗菌肽2、对虾抗菌肽3和对虾抗菌肽4不同。在包括卵巢和下颌器官在内的各种组织中检测到了mo-penaeidin mRNA。mo-penaeidin mRNA在从单细胞期到幼体后期阶段均有存在,在无节幼体I期(孵化后9小时)水平较高,并且在蜕皮间期表达较高,表明其具有早期先天性免疫以及在蜕皮期具有不同的免疫反应。