Genotyping of T-DNA inserted transgenic rice plants(Oryza stiva) using genomic DNA(g-DNA) and GUS specific primers by PCR amplification. 식물에서 DNA 추출 및 삽입돌연변이체의 유전자형 선별법

소개

T-DNA에 Gus라는 report gene이 사용되어 T-DNA가 벼에 삽입되어 나타나는 genotype을 구분하는 방법에 대해 기술하였다. gene에 대한 isolation과 이 실험 과정에 대한 PCR, electrophoresis에 대해 소개하고 genotyping에 대한 실험 방법과 결론, 고찰에 대해 기술하였다. primer를 만들 때 필요한 간단한 방식에 대해 언급하고 다른 primer를 만들때 필요한 site 및 primer를 적어 놓았다. 논문 형식에 따른 영문 reporte로써 필요하신 분들은 참조하세요

목차

ABSTRACT: 실험에 대한 요약
Key words : 실험에서 중요한 단어들
INTRODUCTION : T-DNA, Isolation genomic DNA, PCR(Polymerase chain reaction), Electrophoresis 에 대한 소개글들.
MATERIALS & SUPPLIES : 실험에서 사용한 기구 및 시약
PROCEDURE : 실험과정
RESULT : 실험 결과
DISCUSSION : 이번 실험에 대한 고찰
CONCLUSION : 실험에 대한 총제적인 결과
Futher Reading : 더 읽어 볼 논문.
REFERENCE : 참조글

본문내용

INTRODUCTION
T-DNA(transfer DNA) that is the transferred DNA located the tumor-inducing (Ti) plasmid of some species of bacteria such as Agrobacterium tumefaciens and Agrobacterium rhizogenes[Fig 1-1]. It derives its name from the fact that the bacterium transfers this DNA fragment into the host plant`s nuclear DNA genome using RB and LB[Fig 1-2]. grobacterium-mediated T-DNA transfer is widely used as a tool in biotechnology. In genetic engineering, the tumor-promoting and opine-synthesis genes are removed from the T-DNA and replaced with a gene of interest or a selection marker. Agrobacterium is then used as a vector to transfer the engineered.
T-DNA into the plant cells where it integrates into the plant genome. This method can be used to generate transgenic plants carrying a foreign gene(=2.5).
This experiment is finding out rice plant`s genotype inserted T-DNA that contains inserted gene called GUS(beta-glucuronidase) located adjacent RB. In experiment, necessary process is isolation genomic DNA, PCR, electrophoresis.
Isolation genomic DNA
The use of DNA for analysis or manipulation usually requires that it be isolated and purified to a certain extent.

참고자료

· 1. David L. Nelson and Michael M. Cox. Lehninger Principles of biochemistry 4th edition. Freeman 2005; p283-284
· 2. http://insilico.ehu.es/tm.php
· 3. Keith wilson&John Walker. Principles and Techniques of biochemistry and molecular biology 6th edition. Cambridge 2005; p191-192, 207-209, 449-476
· 4. http://en.wikipedia.org/wiki/T-DNA
· 5. Lincoln Taiz & Eduardo Zeiger. Photosynthesis: Cytokinins. In: Plant physiology 2th edition. Sinauer Associates. 1998; p627-628