A Study On The Related Genes Of Early Embryonic Development Of Chinese Artemia Parthenogenetica

The brine shrimp Artemia is normally restricted to saline inland lakes and coastal salterns. It belongs to Crustasea, Branchiopoda, Anostacea, Artemidae, Artemia. In this experiment, the related genes of early embryonic development of Chinese Artemia Pathenogenatica have been studied by mRNA Differential Display. RNA samples, extracted from six developmental phases: Oh, 4h, 8h, 12h, 16h, 10d, were used as templates. DDRT-PCR was performed with three anchored primers for cDNA synthesis and 26 random primers in the PCR. After running on 6% non-denaturing PAGE, the differential expressed DNA bands were stained by Silver Nitrate. 286 DNA fragments related to the Genes of Early Embryonic Development were isolated. The size of these fragments was 250~2000 bp in length. 24 DNA fragments were cloned into vector and sequenced. The results were analyzed by BLAST through GeneBank/EMBL/DDBJ databases. Seql and Seq2 DNA fragments showed high sequencial homology to known genes. The other fragments showed low degree of similarity to previously reported genes. Seql showed high homology to Oreochromis mossambicus muscle-type creatine kinase CKM2 mRNA, and the part region identity with the sequence of CKM2 mRNA is 92% (482/517), Score=747, E=0. The BLASTX procedure was used to analyze Seql. The result illuminated that the amino acid sequences deduced from Seql showed high degree of similarity to Oreochromis mossambicus muscle-type creatine kinase CKM2, the part region identity with the sequence of Oreochromismossambicus muscle-type creatine kinase CKM2 is 88%(147/167), Score = 300 bits (700), Expect (2) = 3e-80. On the stages of 16h and lOd the gene was expressed more than that of the other stages on the PAGE. RT-PCR and sequencing analysis indicated that it was really expressed in 16h and lOd. Seql is a fragment of the CKM2 gene of Chinese Artemia Parthogenetica, and the gene is related to early embryonic development of Chinese Artemia Parthenogenetica. Seq2 showed high homology to Caenorhabditis elegans seryl tRNA Synthetase by BLASTX procedure analysis, and the part region identity with the amino sequence of tRNA Synthetase is 55%(55/100), Score=113, E=2e—24. The definite functions of the two genes need further study. The fragments showing low degree of similarity to previously reported genes may be new genes. The further research on these sequences is very useful to obtain the complete genes and to know the mechanism of early embryonic development of Chinese Artemia Parthenogenetica.