RAPD Analysis On Genomic DNA Of Two Artificial Gynogenetic Groups Of Grass Carp And The Genomic Structure Analysis Of Goldfish Vsx1 Gene

Part 1: RAPD analysis on genomic DNA of two artificial gynogenetic groups of grass carp, Ctenopharyngodon idellu.Genetic similarity, genetic distance and Shannon's genetic diversity of two different artificial gynogenetic grass carp groups and a control common grass carp group were analyzed with the methods of RAPD (Random Amplified Polymorphism DNA). One of the two different gynogenetic groups was a meiotic-mito-gynogenetic grass carp group that was produced by artificial gynogenesis for two generations. Another gynogenetic group was a mito-gynogenetic grass carp that was generated by artificial gynogenesis only one generation. The common grass carp group was randomly collected from the markets of Changsha.Total 291 loci were detected in all of the three groups through the RAPD-PCR with 26 polymorphic random primers. The loci number in the three groups, meio-mito-gynogenetic group, the mito-gynogenetic group and the common grass carp group, were 265, 272 and 282 and the number of polymorphic loci were 15, 19 and 81 respectively. Genetic statistical analysis showed that percentage of polymorphic loci of the three groups of grass carp, were 5.66%, 6.99%, 28.23%; and the genetic similarity within the three groups of grass carp estimated by Nei's index were 0.9851, 0.9820, 0.9114, respectively. The genetic diversity indices within the three groups of grass carp estimated by Shannon index of phenotypic diversity were 0.1722, 0.3169 and 0.8450 respectively. Generally, the genetic diversity within two artificial gynogenetic groups of grass carp is far lower than that of common grass carp, while thegenetic diversity in meiotic-mito-gynogenetic grass carp group is much lower than that of mito-gynogenetic grass carp group. The relatively low level of genetic diversity in each of the two groups of gynogenetic grass carp revealed that the population purity in each of the two gynogenetic grass carp groups is higher than that of common grass carp group.Part 2: The genomic structure analysis of goldfish vsxlvsxl is a gene encoding a protein with a paired-kile homeodomain and a CVC domain. It is related with early development of goldfish' s eye and plays an important role in the proliferation of retinal progenitor cell, specification and differentiation or maintenance of rod and cone bipolar cells. In this study, we isolated and cloned the 5' flanking sequence and genomic sequence of goldfish Vsxl gene, which provides the foundation for exploring the function and expressing mechanism of vsxl.Three nested gene specifical primers were designed according to the known partial 5' sequence of goldfish vsxl. During the course of TAIL-PCR, the three primers are combined with one of totally 11 AD (arbitrary degenerate). DNA extracted from goldfish fin was used as template of TAIL-PCR reaction. Three Fragments which were denominated Fa3, Fb3, Fd3 respectively were obtained through TAIL-PCR, then the three Fragments that were purified were sequenced. The sequencing results show that the fragment Fd3 is different from Fa3 and Fb3, among these upstream sequences of transcriptal start site of vsxl, TATA box, namely Hogness box, has not been found.The primer set of IS and 2A that were designed according the goldfish vsxl were used to amplify the genomic sequence of vsxl...