| A wide variety of sources, biodegradable, environmentally friendly high calorific value and other advantages of biodiesel has become a hot new energy research various countries of the world. Microalgae have not occupied the land, easy to train, and high fat content. It is considered the best raw material for biodiesel. This study show the expression of LPAAT gene and the content of fatty acid with the methods such as the fluorescent quanfitation, GC-MS, and laser confocal microscopy by transferring over-expression vector and anti-sense expression vector into Phaeodactylutum tricornutum with the method of microprojectile bombardment.Here, we reported that the Over-expression vector LPAAT / Bar and antisense expression vector anti-LPAAT / Bar was constructed successfully. two recombinant vectors were successfully transferred within the Phaeodactylum tricornutum by biolistic method. LPAAT gene expression of the transformants B of overexpression group was maximum and the transformants F was lowest.The results of fatty acid analysis indicate that in the logarithmic phase, the total fat of the over-expression group is 1.86 times that of the control group, the total fat of the anti-sense group is 26% of that of the control group, and the total fat of the over-expression group and the anti-sense group differs significantly from that of the control group(p<0.01). At the beginning of the plateau, the total fat of the over-expression group is 1.24 times that of the control group, the total fat of the anti-sense group is 26% of that of the control group, the total fat of the over-expression group differs significantly from that of the control group(p<0.05), and the total fat of the anti-sense group is 66% of that of the control group, so the total fat of the anti-sense group differs significantly from that of the control group(p<0.05). At the end of the plateau, the total fat of the over-expression group is 1.4 times that of the control group, which differs significantly from that of the control group(p<0.05). The total fat of the anti-sense group is 85% of that of the control group, so the total fat of the anti-sense groupdiffers only slightly from that of the control group. Under the same conditions the red fluorescence in the control group, the antisense group and overexpression group is basically. The results show Nile Red into the algal cells are basically the same, but the yellow fluorescence of the over-expression is approximately 1.3 times to control group, yellow fluorescence of antisense group was 63% to the control group. suggesting that with yellow fluorescence in the overexpression group are more fat contents, and in the antisense group LPAAT gene expression was inhibitted. So the fat content was less. The LPAAT gene of the preference of C18 fatty acids, over-expression group compared with the control group, in the logarithmic phase, initial stage platform and final stage platform C18:2>C18:1>C18: 3.In logarithmic phase, the relative expression quantity of LPAAT gene of the over-expression group is about 2.4 times higher than that of the control group, so the relative expression quantity of LPAAT gene differs significantly between the over-expression group and the control group. The relative expression quantity of LPAAT gene of the anti-sense group is 69 % that of the control group,so the relative expression quantity of LPAAT gene of the anti-sense group differs significantly from that of the control group(P<0.01). At the beginning of the plateau, the relative expression quantity of LPAAT gene of the over-expression group is about 1.3 times higher than that of the control group, so the relative expression quantity of LPAAT gene of the over-expression group differs significantly from that of the control group. The relative expression quantity of LPAAT gene of the anti-sense group is 48 % that of the control group,so the relative expression quantity of LPAAT gene of the anti-sense group differs significantly from that of the control group(P<0.05). At the end of the plateau, the relative expression quantity of LPAAT gene of the over-expression group is about 1.6 times higher than that of the control group, so the relative expression quantity of LPAAT gene of the over-expression group differs significantly from that of the control group. The relative expression quantity of LPAAT gene of the anti-sense group is 40% that of the control group,so the relative expression quantity of LPAAT gene of the anti-sense group differs significantly from that of the control group(P<0.01).Because siliceous shell of Phaeodactylum, traditional transformation method is difficult to transformed vector into Phaeodactylum cells. In this study, by the gene gun transformation method, transformed overexpression vector and inhibition of gene expression vector ofLPAAT Successful. The overexpression and inhibition of gene expression LPAAT, over ability to express algae strains greatly increased accumulation of fatty acids, fatty acids antisense algae strains restrained, this result is the development of microalgae biodiesel industry,it provides a theoretical basis for bioenergy... |
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