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Isolation of Carotenoid Genes from Lycium Barbarm

Author: ZhangJi
Tutor: JiJing
School: Tianjin University
Course: Biochemical Engineering
Keywords: lycium barbarm cDNA library DIG-DNA labeling ZDS gene DIG
CLC: Q943
Type: Master's thesis
Year: 2007
Downloads: 205
Quote: 0
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Abstract


Carotenoid is a kind of pigment which is wild existent in the nature. Carotenoids are a group of phytochemicals that have strong antioxidant properties and therefore protect biological structures, so it plays an important role in the nutrition of human. At present, people have a great interest on the effect of carotenoid in improving the crop’s nutrition, improving appearance character and changing the design and color of ornamental.RNA extraction plays an important role in the construction of cDNA library. The operation of RNA includes purifying RNA, amplifying via RT-PCR and so on. In the experiment, aimed at the existence of disturb composition, getting the suitable extraction method. This makes a well groundwork for the building of cDNA library.Using the root, stem and leaf of lycium barbarm as the stuff, with contrast of TRIZOL Reagent Method, CTAB-acidic Phenolic Method, isothiocyanate method and SDS- phenol Method, the ideal method for extraction of total RNA from the root, stem and leaf of lycium barbarm was established. In the experiment, overcome the hardness that the RNA form the root is very hard to be extracted, and getted good quality RNA. At the same time, assisted building the lycium barbarm cDNA library and the capacity of the library was 2.78×10~5pfu, and the recombination rate reached to 88.1%.Heterologous probe of PDS、ZDS originate from Getina lutea were labeled with digoxingenin-11-dUTP (DIG), were further used to screen the cDNA libraries of leaf of lycium barbarm. Two ZDS positive clones were obtained.This study will lay foundations for regulating carotenoid biosynthesis via genetic engineening.

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