首次在长牡蛎(Crassostrea gigas)中克隆得到一种新的贝壳基质蛋白nacrein-like protein F3的全长c DNA序列。nacrein-like protein F3基因c DNA全长1499bp,其中编码区长度为1242bp,编码一条含413个氨基酸残基的多肽链。氨基酸序列比对和结构域分析均表明其为合浦珠母贝(Pinctada fucata)nacrein的同源蛋白,含有1个保守的α-碳酸酐酶结构域,但由于重复结构域的插入,α-碳酸酐酶结构域被间隔成2个亚结构域。系统进化分析显示nacrein-like protein F3与贻贝(Mytilus californianus)nacrein-like protein进化关系最近。此外,在软体动物中,双壳纲nacrein-like proteins进化速度相对较快,推测与寒武纪时期剧烈的环境变化有关,如影响贝壳形成的海水化学变化。
Microsatellites are a ubiquitous component of the eukaryote genome and constitute one of the most popular sources of molecular markers for genetic studies. However, no data are currently available regarding microsatellites across the entire genome in oysters, despite their importance to the aquaculture industry. We present the fi rst genome-wide investigation of microsatellites in the Pacifi c oyster Crassostrea gigas by analysis of the complete genome, resequencing, and expression data. The Pacifi c oyster genome is rich in microsatellites. A total of 604 653 repeats were identifi ed, in average of one locus per 815 base pairs(bp). A total of 12 836 genes had coding repeats, and 7 332 were expressed normally, including genes with a wide range of molecular functions. Compared with 20 different species of animals, microsatellites in the oyster genome typically exhibited 1) an intermediate overall frequency; 2) relatively uniform contents of(A)n and(C)n repeats and abundant long(C)n repeats(≥24 bp); 3) large average length of(AG)n repeats; and 4) scarcity of trinucleotide repeats. The microsatellite-fl anking regions exhibited a high degree of polymorphism with a heterozygosity rate of around 2.0%, but there was no correlation between heterozygosity and microsatellite abundance. A total of 19 462 polymorphic microsatellites were discovered, and dinucleotide repeats were the most active, with over 26% of loci found to harbor allelic variations. In all, 7 451 loci with high potential for marker development were identifi ed. Better knowledge of the microsatellites in the oyster genome will provide information for the future design of a wide range of molecular markers and contribute to further advancements in the fi eld of oyster genetics, particularly for molecular-based selection and breeding.
Fibrinogen-related proteins(FREPs) are lectins with at least a fibrinogen-like domain.They are widespread in Mollusca and play an important role as immune pattern-recognition receptors.These diverse molecules provide interesting models for the study of the evolution of innate immunity.Although oysters are among the most studied mollusks,there have been few investigations of this gene family in oysters.This paper reports the cloning of the first oyster FREP,named CgFREP,from the Pacific oyster Crassostrea gigas.Five types of FREP(CgFREP-1 to CgFREP-5) were obtained based on a single pair of primers,confirming their high diversity.The proteins deduced from these FREPs contained 253-363 amino acids and were divided into three subfamilies based on sequence identities(cutoff point <86%).The topological structure of the phylogenetic tree showed the five CgFREPs clustered together and were completely separated from the subgroup of FREPs obtained from Biomphalaria glabrata(BgFREPs).This implies that FREP diversity evolved before the BgFREPs branched off.An interesting aspect of the CgFREPs was their individual hypermutation.Ninety-three SNPs and five indels were characterized among CgFREPs.Some mutations(e.g.an insertion in CgFREP-5) could cause premature translation termination.Multiple alignments suggest recombination events might have occurred during exchanges between alleles,or by somatic diversification,or both.Individual sequence variability parallels the extraordinary complexity of the innate immune system,suggesting a potential capacity to recognize and eliminate many kinds of pathogens,even in a primitive system.The diversity of immune pattern-recognition receptors may offer an alternative strategy that functions in the absence of highly specific immune globulins,which only emerged in vertebrates.
Microsatellites were screened in a backcross family of the Pacific oyster, Crassostrea gigas. Fifteen microsatellite loci were distinguishable and polymorphic with 6 types of allele-combinations. Null alleles were detected in 46.7% of loci, accounting for 11.7% of the total alleles. Four loci did not segregate in Mendelian Ratios. Three linkage groups were identified among 7 of the 15 segregating loci. Fluorescence-based automated capillary electrophoresis (ABI 310 Genetic Analyzer) that used to detect the microsatellite loci, has been proved a fast, precise, and reliable method in microsatellite genotyping.
