The Studies On Cloning And Analysis Expression Of DMRT2Gene In Amphioxus Branchiostoma Belcheri Tsingtauense

The DMRT gene family is a gene family related to sexual determination. Transcriptionfactors encoded by DMRT have a conserved DM-domain linking to DNA by special zincfinger structures. The present studies of DMRT genes show that the DMRT gene familymainly participate in sexual differentiation of embryo, developing of gonads, formation ofgametes, and take part in other functions such as neurodevelopment. This thesis mainlystudied the cDNA and DNA base sequence of DMRT2gene, and studied the expression ofDMRT2gene.Amphioxus is chordata and is belong to cephalochordate, as transitional type it connect thevertebrate and invertebrate, and it’s the vertebrate who most similar to invertebrate.Becausethe amphioxus have the features of vertebrate’s form and the development model, and it’sgenome is simple, it often used be model organism to study the origin of vertebrate and theevolution of invertebrate. Therefore, it is meaningful to study the sexual differentiation ofamphioxus, and the DMRT gene family as an important member related to sexualdifferentiation and developing of gonads, the study of it become instructive and meaningful.This thesis on the basis of the conservative DM domain of the DMRT gene family,through blast the DMRT2sequence of closely related species to amphioxus, then get thecDNA sequence of amphioxus, by means of RACE(Rapid amplification of cDNA ends)to getthe total base sequence of DMRT2gene. Based on the1550bp cDNA sequence, we get theDNA base sequence451bp, then by means of genome walking, we get2210bp DNAsequence of DMRT2gene. After the gene sequencing, we by means of ISH(In situhybridization) to study the expression of the DMRT2gene, we find that DMRT2isexpressing in gonad of amphioxus, not in gill and muscle tissue. All of above is providing ameaningful reference to the study of sexual differentiation and determination of Amphioxus.