江西农业学报
江西農業學報
강서농업학보
ACTA AGRICULTURAE JIANGXI
2015年
5期
65-70
,共6页
段庆虎%张应香%龚凤萍%竹玮%韩玉玲%尹川川%祁玉良%段仁周
段慶虎%張應香%龔鳳萍%竹瑋%韓玉玲%尹川川%祁玉良%段仁週
단경호%장응향%공봉평%죽위%한옥령%윤천천%기옥량%단인주
真菌%漆酶基因%密码子偏好性%密码子优化%异源表达
真菌%漆酶基因%密碼子偏好性%密碼子優化%異源錶達
진균%칠매기인%밀마자편호성%밀마자우화%이원표체
Fungus%Laccase gene%Codon usage bias%Codon optimization%Heterogeneous expression
漆酶是一种含铜的多酚氧化酶,广泛地存在于各种植物和真菌中。研究使用 CodonW和CUSP相关程序对18条真菌漆酶基因CDS序列的密码子使用情况进行分析,比较了真菌漆酶基因与枯草芽孢杆菌、大肠杆菌以及毕赤酵母的密码子偏好性。结果表明,真菌漆酶基因偏好使用以G /C结尾的密码子,29种偏好密码子(相对使用度( RSCU)>1)中偏好性较强的密码子有ATC、CTC、CGC、GTC和TAG ( RSCU≥1.5);聚类分析发现,AY450404.1和GU480806.1、XM_9518462.和XM_001228805.1及XM_001389488.2和XM_001818019.2聚为一类,与系统分类关系一致,其余都不一致;在密码子的使用频率上,真菌漆酶基因的密码子用法与枯草芽孢杆菌、大肠杆菌和毕赤酵母的基因组密码子用法都有不同程度的偏好差异,其中与大肠杆菌的密码子用法差异最小,故大肠杆菌更适合作为真菌漆酶基因的外源表达宿主。如果要在上述几种表达系统中实现真菌漆酶基因的高效表达则需对真菌漆酶基因的部分密码子进行优化改造。
漆酶是一種含銅的多酚氧化酶,廣汎地存在于各種植物和真菌中。研究使用 CodonW和CUSP相關程序對18條真菌漆酶基因CDS序列的密碼子使用情況進行分析,比較瞭真菌漆酶基因與枯草芽孢桿菌、大腸桿菌以及畢赤酵母的密碼子偏好性。結果錶明,真菌漆酶基因偏好使用以G /C結尾的密碼子,29種偏好密碼子(相對使用度( RSCU)>1)中偏好性較彊的密碼子有ATC、CTC、CGC、GTC和TAG ( RSCU≥1.5);聚類分析髮現,AY450404.1和GU480806.1、XM_9518462.和XM_001228805.1及XM_001389488.2和XM_001818019.2聚為一類,與繫統分類關繫一緻,其餘都不一緻;在密碼子的使用頻率上,真菌漆酶基因的密碼子用法與枯草芽孢桿菌、大腸桿菌和畢赤酵母的基因組密碼子用法都有不同程度的偏好差異,其中與大腸桿菌的密碼子用法差異最小,故大腸桿菌更適閤作為真菌漆酶基因的外源錶達宿主。如果要在上述幾種錶達繫統中實現真菌漆酶基因的高效錶達則需對真菌漆酶基因的部分密碼子進行優化改造。
칠매시일충함동적다분양화매,엄범지존재우각충식물화진균중。연구사용 CodonW화CUSP상관정서대18조진균칠매기인CDS서렬적밀마자사용정황진행분석,비교료진균칠매기인여고초아포간균、대장간균이급필적효모적밀마자편호성。결과표명,진균칠매기인편호사용이G /C결미적밀마자,29충편호밀마자(상대사용도( RSCU)>1)중편호성교강적밀마자유ATC、CTC、CGC、GTC화TAG ( RSCU≥1.5);취류분석발현,AY450404.1화GU480806.1、XM_9518462.화XM_001228805.1급XM_001389488.2화XM_001818019.2취위일류,여계통분류관계일치,기여도불일치;재밀마자적사용빈솔상,진균칠매기인적밀마자용법여고초아포간균、대장간균화필적효모적기인조밀마자용법도유불동정도적편호차이,기중여대장간균적밀마자용법차이최소,고대장간균경괄합작위진균칠매기인적외원표체숙주。여과요재상술궤충표체계통중실현진균칠매기인적고효표체칙수대진균칠매기인적부분밀마자진행우화개조。
Laccase is a kind of polyphenol oxidase which contains copper, and widely exists in all kinds of plants and fungi. The coding sequences of 18 laccase genes from fungi were analyzed to identify their codon usage bias by using CodonW and CUSP programs.The codon bias of fungal laccase genes was compared with that of Bacillus subtilis, Escherichia coli and Pichia pastoir s. The results showed that most of fungal laccase genes were bias toward the synonymous codons with G and C at the third codon posi-tion.There were twenty-nine biased codons [ relative synonymous codon usage ( RSCU )>1.0 ] , among them the codons with stronger bias character (RSCU≥1.5) were ATC,CTC , CGC, GTC and TAG.Clustering analysis showed that AY450404.1 and GU480806.1, XM_951846.2 and XM_001228805.1, XM_001389488.2 and XM_001818019.2 were respectively clustered into a group, which agreed with the taxonomic relationships;while the other clustering results were not the same as the taxonomic relation-ships.The difference in codon usage frequency between fungal laccase genes and genome of Escherichia coli was less than that be-tween fungal laccase genes and genome of Bacillus subtilis or Pichia pastoris.Therefore, Escherichia coli expression system might be more suitable for the expression of fungal laccase genes.For realizing the high-efficient expression of fungal laccase genes in the a-bove three expression systems, it is necessary to optimize several codons of fungal laccase genes.