CAJ | 학술논문

[目的]克隆高羊茅 FaGF14-B和FaGF14-C基因并进行基因的差异表达分析，为后续基因功能研究提供依据。[方法]以高羊茅转录组学测序获得的 FaGF14-B 和FaGF14-C基因序列片段为模板，利用3'RACE和5'RACE方法从高羊茅中扩增出 FaGF14-B和FaGF14-C基因的全长 cDNA序列，命名为FaGF14-B和 FaGF14-C，并进行核酸序列分析、基因编码蛋白分析、蛋白保守结构域分析、系统进化树分析与差异表达分析。[结果] FaGF14-B基因序列 cDNA全长1548 bp，具有完整的开放阅读框（ORF，449~1228 bp），编码蛋白为260个氨基酸；FaGF14-C基因序列 cDNA全长1250 bp，具有完整的开放阅读框（ORF，66~848 bp），编码蛋白含有261个氨基酸；GF14-B和 GF14-C蛋白都具有典型的14-3-3蛋白结构域，二级结构包含9个保守的α-螺旋及不保守的 N-末端和 C-末端。系统进化树分析表明，高羊茅 FaGF14-B 和FaGF14-C与禾本科植物 GF14蛋白具有较高的相似性，位于在同一个进化支上，亲缘关系较近；荧光定量 PCR分析表明：FaGF14-B和FaGF14-C基因应答氮胁迫处理。[结论]该研究可为进一步筛选抗低氮胁迫相关基因、创制耐低氮的牧草新种质奠定理论基础。
[목적]극륭고양모 FaGF14-B화FaGF14-C기인병진행기인적차이표체분석，위후속기인공능연구제공의거。[방법]이고양모전록조학측서획득적 FaGF14-B 화FaGF14-C기인서렬편단위모판，이용3'RACE화5'RACE방법종고양모중확증출 FaGF14-B화FaGF14-C기인적전장 cDNA서렬，명명위FaGF14-B화 FaGF14-C，병진행핵산서렬분석、기인편마단백분석、단백보수결구역분석、계통진화수분석여차이표체분석。[결과] FaGF14-B기인서렬 cDNA전장1548 bp，구유완정적개방열독광（ORF，449~1228 bp），편마단백위260개안기산；FaGF14-C기인서렬 cDNA전장1250 bp，구유완정적개방열독광（ORF，66~848 bp），편마단백함유261개안기산；GF14-B화 GF14-C단백도구유전형적14-3-3단백결구역，이급결구포함9개보수적α-라선급불보수적 N-말단화 C-말단。계통진화수분석표명，고양모 FaGF14-B 화FaGF14-C여화본과식물 GF14단백구유교고적상사성，위우재동일개진화지상，친연관계교근；형광정량 PCR분석표명：FaGF14-B화FaGF14-C기인응답담협박처리。[결론]해연구가위진일보사선항저담협박상관기인、창제내저담적목초신충질전정이론기출。
Objective] The differential expression analysis was performed for FaGF14-B and FaGF14-C genes in tal fescue so as to provide certain basis for fol ow-up functional analysis of genes. [Method] The sequence fragments of FaGF14-B and FaGF14-C obtained from reverse transcription were used as templates, and the ful-length cDNA sequences of FaGF14-B and FaGF14-C were amplified using the 5’ RACE use 3’RACE techniques. They were named as FaGF14-B and FaGF14-C, and used for nucleic acid sequence analysis, encoded protein analysis, protein con-served domain analysis, phylogenetic analysis and differential expression analysis. [Result] The FaGF14-B gene has a ful length of 1 548 bp. It has a complete open reading frame (ORF, 449-1 228 bp), and encodes a protein composed of 261 amino acids. The FaGF14-C gene has a ful length of 1 250 bp. It also has a complete open reading frame (ORF, 66-848 bp), and encodes a protein composed of 261 amino acids. The GF14-B and GF14-C proteins al have a typical domain 14-3-3, and their secondary structures al contain 9 conserved α-helical structures and non-conserved N- and C- terminals. The phylogenetic analysis showed that the FaGF14-B and FaGF14-C from tal fescue have high similarities with GF14 protein from gramineous plants, and they are divided into the same clade with closer ge-netic relationship. The real-time fluorescence quantitative PCR analysis showed that the expression of FaGF14-B and FaGF14-C genes is al sensitive to nitrogen stress. [Conclusion] This study wil lay a theoretical basis for further screening of low nitrogen-tolerant genes and breeding of low nitrogen-tolerant grass germplasms.