Aequorea taiwanensis n. sp. (Hydrozoa, Leptomedusae) and mtCOI sequence analysis for the genus Aequorea

Aequorea taiwanensis, a new hydrozoan species from the Taiwan Strait was described using morphological and molecular characteristics. Both morphological and mitochondrial cytochrome oxidase subunit I (mtCOI) data supported A. taiwanensis n. sp. as a valid species. Sequence divergence and genetic distance of A. taiwanensis n. sp., A. papillata and A. conica were analysed based on the mtCOI gene sequences. The mtCOI sequences from these three species of the genus Aequorea showed high variation frequency, with sequence divergences ranging from 9.10% to 11.9%, and pairwise genetic distances ranging from 0.097 to 0.130. MtCOI sequence analysis provided diagnostic molecular systematic characteristics for accurate identification and discrimination of the species of Aequorea or their populations, and will be used to resolve evolutionary relationships among them.It was suggested that 10%—20% pairwise mtCOI sequence differences indicated the specieslevel divergence among congeneric species in the Hydromedusae.     

二种多管水母光蛋白基因的分离、表达及生物活性初步研究

分别从厦门东海域的大型多管水母和细小多管水母中分离到了新的光蛋白基因aeqxm和aeqxxm,并在大肠杆菌中进行了表达.aeqxm和aeqxxmDNA序列的编码区总长均为585bp,无内含子序列,推导的氨基酸序列总长均为195个氨基酸.aeqxm,aeqxxmDNA和AEVAQ440XcDNA之间的核苷酸序列同源性分别为80.7%,85.1%,aeqxm和aeqxxmDNA的核苷酸序列同源性为87.2%.原光蛋白apoaeqxm,apoaeqxxm与AEVAQ440X编码的氨基酸序列之间的同源性分别为84.7%,84.2%,apoaeqxm和apoaeqxxm的氨基酸序列之间的同源性为94.4%.分别将aeqxm和aeqxxm基因克隆至pTO-T7表达载体,apoaeqxm,apoaeqxxm在大肠杆菌中的表达量都达到40%左右.取菌体超声上清与腔肠动物荧光素f、巯基乙醇混合再生后,加入CaC_l2,用荧光全谱仪检测到470nm处的瞬时发光,表明表达的apoaeqxm,apoaeqxxm具有正常的生物学功能.     

大型多管水母的绿色荧光蛋白

采用PCR扩增和southern杂交筛选相结合的方法,从厦门水域的大型多管水母中分离到了新的绿色荧光蛋白基因gfpxm,并在大肠杆菌中进行了表达.gfpxmDNA序列编码区长为1042bp,包含3个外显子和两个内含子,cDNA编码区全长为717bp.gfpxmcDNA与已知野生型Aeqgfp 10cDNA长度一致,核苷酸同源性为81.9%,推导的氨基酸序列全长为238个氨基酸,与AeqGFP10的氨基酸同源性为83.6%.将gfpxm克隆至pTO-T7表达载体,GFPxm在大肠杆菌BL21中的表达量达菌体总蛋白的50%左右.荧光性质和强度分析结果表明,GFPxm蛋白的激发峰为476nm,发射峰为496nm,荧光量子产率为1. GFPxm蛋白的荧光很稳定,对热、碱性、变性剂和盐等有较强抗性.     

Cloning and expression of aequorin genes from jellyfish Aequorea and characterization of aequorins activities

ＩＮＴＲＯＤＵＣＴＩＯＮＡｅｑｕｏｒｉｎｉｓａｂｉｏｌｕｍｉｎｅｓｃｅｎｔｐｒｏｔｅｉｎ ,ｉｓｏｌａｔｅｄｆｒｏｍ ｊｅｌｌｙｆｉｓｈＡｅｑｕｏｒｅａ .Ｔｈｅａｅｑｕｏｒｉｎｃｏｍｐｌｅｘｃｏｎｓｉｓｔｏｆａ 2 2 0 0 0Ｍｒａｐｏａｅｑｕｏｒｉｎｐｒｏｔｅｉｎ ,ｍｏｌｅｃｕｌａｒｏｘｙｇｅｎａｎｄｔｈｅｌｕｍｉｎｏｐｈｏｒｅｃｏｅｌｅｎｔｅｒａｚｉｎｅ .ＷｈｅｎｔｈｒｅｅＣａｉｏｎｓｂｉｎｄｔｏｔｈｉｓｃｏｍｐｌｅｘ ,ｃｏｅｌｅｎｔｅｒａｚｉｎｅｉｓｏｘｉｄｉｚｅｄｔｏｃｏｅｌｅｎ ｔｅｒａｍｉｄｅ…     

2011年春季黄、东海墨绿多管水母(Aequorea coerulescens)分布特征

2011年4月在黄、东海的大面调查中观测到一种多管水母大量发生,在现场通过表层目测计数了其分布特征,采集现场样品测量了伞径、湿重和干重,并结合水文数据分析了其分布与水团的关系。经鉴定,该种为水螅水母纲、软水母亚纲、锥螅水母目、多管水母科、多管水母属、墨绿多管水母(Aequorea coerulescens)。在69个站位中,有11个站位出现,出现率16%,主要在东海南部近岸海域。个体伞径在17.40—142.00mm之间,湿、干重分别在3.07—75.47g和0.12—3.06g之间,湿、干重与伞径呈现显著的指数关系。碳含量占干重的3.15%±0.56%,氮含量占干重的14.44%±2.65%,碳氮比为4.58±0.30。伞径和温度、盐度显著相关。作者认为,墨绿多管水母来源于舟山群岛附近海域,受海流的作用,输送到观测海域;离源地越远的站位,由于生长时间长,水母伞径增大。