Scge And Rapd Analysis Of DNA Damage Effects Induced By Petroleum Hydrocarbons Of Hemicentrotus Pulcherrimus

There are more and more accidents of petroleum hydrocarbon leaks in the world today, causing serious pollution to the marine ecological environment.This paper takes the Hemicentrotus pulcherrimus as the test object, using SCGE and RAPD to analysis the damage which is caused by petroleum hydrocarbon to sea urchin at the cellular level and the molecular level,exploring the feasibility of these two technologies in the detection of marine petroleum hydrocarbon contamination, providing a reference for early warning and risk assessment of marine environmental pollution.Taking 0# diesel gradient concentration and exposure days as the variable conditions, study the damage which is caused by petroleum hydrocarbon in the level of cells with SCGE. The result shows that the DNA of the Hemicentrotus pulcherrimus appears the phenomenon of Comets trailing when the 0#diesel concentration is 50 mg/L. So, the result shows that the DNA of the Hemicentrotus pulcherrimus will be damaged when the 0#diesel concentration is 50 mg/L. This paper shows the relationship of time-effect between exposure days and cell damage rate, tail DNA percentage (%DNAT) and the average moving length of DNA when the 0#diesel concentration is 50 mg/L.Taking 0#diesel gradient concentration and exposure days as the variable conditions, study the damage which is caused by petroleum hydrocarbon in the level of molecular with RAPD. In order to find the best reaction system, to filter out the random primers which can amplify the best effect and to make the experimental effect to be more obvious.Firstly, the writer use RAPD to analyze the genetic diversity of the Hemicentrotus pulcherrimus which haven’t been poisoned. The result shows that the optimal conditions of RAPD-PCR system for Hemicentrotus pulcherrimus will be performed in a 25μL reaction mixture with 0.2μL Taq enzyme,2.5μL 10×Buffer,2μL dNTP,1μL template DNA,1μL random primer,18.3μL ddH20. There are 15 random primers that have been sifted for doing the PCR amplification of genomic DNA fragment. There are 504 bands total, averaging 19 bands of each one.75.4% of the bands are polymorphic. The genetic distance between individuals is 0.2189-0.5307, and the average genetic distance is 0.3254. So, the genetic diversity of Hemicentrotus pulcherrimuses is good. Then put the Hemicentrotus pulcherrimuses in the 0#diesel solution, studying the damage of Hemicentrotus pulcherrimuses DNA by RAPD. The result shows that only when the 0# diesel concentration is 50 mg/L, the shading value of the bands is changed, especially when the primers are S49 and S67. So, the petroleum hydrocarbon can cause the damage of Hemicentrotus pulcherrimuses DNA. And the DNA has easy damaged sites. Because the time that polymorphic bands become dark is different, so the easy damaged sites have different stain resistance.