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Single Nucleotide Polymorphism Analysis of Acetyl-CoA Carboxylase Gene in Brassica Napus. L

Author: RenQiang
Tutor: LiGuanRong；LiJiaNa
School: Southwestern University
Course: Biochemistry and Molecular Biology
Keywords: Brassica napus Acetyl coenzyme A carboxylase (ACCase) Single nucleotide polymorphism (SNP), the oil content of
CLC: S565.4
Type: Master's thesis
Year: 2007
Downloads: 199
Quote: 1
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Abstract

Acetyl coenzyme A carboxylase (ACCase), a biotin prosthetic group, HCO ₃ ^{- carboxylase carboxyl donor need Mg ^{2 participate in the formation of malonyl coenzyme A in vivo catalytic ATP-dependent carboxylation of acetyl-CoA. ACCase catalyzed by the fatty acid synthesis pathway, this step is the rate limiting step in fatty acid synthesis. ACCase is a key enzyme to control the rate of fatty acid synthesis plants, control of the synthesis of fatty acid in the amount of enzyme and the enzyme activity on two levels is the one of the key enzymes of the plant survival. Rape is one of the world's top five oil crops, and is also one of the most important oil crops in China, occupies an important position in China's agricultural production. Single nucleotide polymorphisms (Single nucleotide polymorphism, SNP) refers to the variation of a single nucleotide in the genome, because it has a large number of widely distributed, stability, easy typing advantages, is considered following the restriction fragment length polymorphism (RFLP), amplified fragment length polymorphism (AFLP), simple repeat sequence (SSR) after the fourth-generation of molecular markers. Brassica napus due to the high oil content of the good traits more recently, the importance of Brassica napus (Brassica napus L.) acetyl coenzyme A carboxylase gene polymorphism and the functional relationship is unclear. Study of rape acetyl coenzyme A carboxylase gene can fill this lack of research, to help further research the Brassica napus acetyl coenzyme A carboxylase gene structure and function relationship also establish related genes of Brassica napus oil content SNP molecular markers to provide information. This study used 12 parts of a different oil content selfing homozygous based material, wherein the high oil content (42.91 to 49.75%) and the material of the low oil content (22.5 ~ 25.9%) of 6 parts. Sub-cloned ACCase gene sequencing and its single nucleotide polymorphism analysis and its findings are as follows: 1. Brassica napus ACCase gene cloned according to the the homogeneous type ACCase gene sequence accession number AJ13866 Brassica napus, is divided into nine sections, nine pairs of overlapping sets of primers, and amplified fragment length 800-1200bp sub-cloning 12 materials ACCase gene, which contains most of the coding sequence. 12 parts of the material have been efficient and specific amplification. Obtained by cloning and sequencing, a the ACCase gene sequences of the respective materials. 2. The Brassica napus ACCase gene single nucleotide polymorphism amplified fragment bidirectional sequencing, sequencing results DNASTAR software CLUSTAL analysis, analysis the results with DNASP software polymorphisms parameter calculation. Total 88824bp genomic nucleotide sequence tested materials were detected 317 SNP frequency of occurrence of 1/279; SNP in the coding region of 179, a non-coding region of 138; ACCase gene coding region variation small in the non-coding region; the nonsynonymous mutations Mean (Ka) and synonymous mutations Mean (Ks) ratio is less than 1, indicating that the gene is more conservative. 3. The relationship of Brassica napus the ACCase gene single nucleotide polymorphisms and oil content traits found in the high oil content material with a low oil content of the material is significantly different from the mutation hotspot, located at 5000 ~ 6500bp (ie, the first full genome sequence between 9088 to 10588bp), the mutation hotspot contains four exons and three introns, the region encoding homogeneity 989-1384 ACCase sequence of 396 amino acids, this sequence is located BCCP between CT functional domains, the function of this region is unclear, speculated that this regional variation may improve ACCase activity, resulting in increased oil content in the seeds. The correlation between oil content traits such ACCase gene single nucleotide polymorphisms and Brassica napus, has yet to be confirmed by further studies, in order to draw conclusions.}}

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