Sequence Analysis And Expressions In Different Development Stages Of Trh Gene & APH-1 Gene Of Artemia Sinica

Artemia sinica belongs to Phylum Arthropoda, Class Crustacea, Subclass Branchiopoda, Order Anostaca, Family Artemiidae and Genus Artemia. It lives in the high-salt salt lakes and salterns which exist in Shanxi, Tibet, Xinjiang, Qinghai and Inner Mongolia. So, osmoregulation plays an important part in keeping its life. In the different stage of development, osmoregulation is accomplished by different tissues. For example, when Artemia is hatched into nauplius, it is accomplished by salt gland and thoracic epipods shows its affect in the sub-adult and adult stages. Following the mRNA sequence of A.franciscana trh gene on GenBank, the article uses total RNA of Artemia sinica as template and gets 782bp-long gene fragment by RT-PCR. After the analysis on presumptive amino acid sequence and related protein amino acid sequence supporting by Blastn, software ClustalX 1.83 and MEGA 3.0 software, we demonstrated that this sequence is the part of trh gene of Artemia sinica and its consanguinity is the closest with A.franciscana. Making use of this segment, we design the probe of RNA, and then make WFISH to the Artemia sinica of different development stages. We found that in the period of pre-naupliu, nauplius, latter-nauplius, trh gene expresses in the salt gland of Artemia sinica. In the same time, lightly expresses in the anlages of thoracic epipod of latter-nauplius. However, in the period of sub-adult and adult, it expresses in the thoracic epipods of Artemia sinica. We deduced that the trh gene is an important factor of the osmoregulation of Artemia sinica. Basing on the mRNA sequence of A.franciscana APH-1 gene in the GenBank designing primer, taking the total RNA as template and through RT-PCR, we gain a sequence whose length is 792bp. After analyzing the assumed amino acid sequence of Artemia sinica APH-1 gene and molecular evolution of close amino acid sequence by using blast contrast, the software Clustal X 1.83 and MEGA3.0, we found that the APH-1 comparability between Artemia sinica and A.franciscana is the most. It is the part of sequence of Artemia sinica APH-1 gene. Making use of this segment, we design the probe, and then make WFISH to the Artemia sinica of different development stages. And then we find in the stage of pre-naupliu, nauplius and latter-nauplius, APH-1 appears in salt gland of Atemia sinica but not in sub-adult and adult, so we think APH-1 may play an important function in osmoregulation and the development of salt gland.