Study Of The Characteristics Of Gene Expression Of Prorocentrum Donghaiense

Although its scientific name is in debate, Prorocentrum donghaiense, one of dinoflagellates, has caused most frequent large-scale red tides in Changjiang River Estuary and Zhoushan Mountain Archipelago in recent years, bringing us tremendous economic losses and local environments serious ecological impacts.Studies of dinoflagellate are of importance for both their biology and our understanding of red tide process. Currently, very rich publications concerning the life history, ecology, taxonomy and morphology of P. donghaiense are available. In contrast, molecular biological studies of P. donghaiense are very scarce.In order to initiate the molecular biological research of P. donghaiense, a cDNA library was constructed for the artificial culture of this species at exponential growth phase. The first batch of 565 usable sequencing reads was generated from 700 clones randomly selected and sequenced. Here our trial identification of functional genes of P. donghaiense from these reads with emphasis on those involved in cell proliferation and death were reported. In addition, mitochondrial cob and cox1 gene fragments of P. donghaiense were cloned and sequenced.In the cDNA library construction, total RNA was extracted by UNIQ-10 Trizol RNA Preparation Kit, and cDNAs were synthesized and amplified by cDNA Synthesis Kit and PCR cDNA Library Kit. Finally, the ligated cDNAs with pMD 18-T vector were transducted into E.coli JM 109. The constructed cDNA library was used to the latter analysis of EST.An appropriate approach of identifying functional genes of a species is to generate and analyze expressed sequence tags (EST). EST analysis has been implemented in functional genomic researches of a wide range of eukaryotes. EST are short cDNA sequences that serve as not only gene tags, but also the basis of multiple uses such as profiling transcripts, making cDNA arrays and most importantly, identifying functional genes. A cDNA library was constructed for P. donghaiense at exponential growth phase, and 565 usable sequencing reads were obtained from 700 clones selected randomly. Messenger RNA corresponding reads were clustered into 36 contigs and 272 singletons (EST groups). Comparisons using BlastX program...