摘要
以野生香蕉叶片为试材,采用RT-PCR结合RACE法,通过T/A克隆测序,获得野生香蕉几丁质酶基因全长序列,并在GenBank登录(登录号为:FJ858155)。野生香蕉几丁质酶基因(ChiⅠ1)全长1115bp,包含924bp的ORF、14bp的5'非编码区、177bp的3'非编码区及17bp3'poly(A)尾。该基因开放阅读框编码307个氨基酸,分子量为32424.1u,预测的理论等电点为6.28,其结构包括信号肽、几丁质结合区、糖苷水解酶区。蛋白结构分析结果表明,该基因编码蛋白为跨膜蛋白,存在于胞外。野生香蕉几丁质酶基因(ChiⅠ1)属于酸性几丁质酶classⅠb,与大蕉几丁质酶classⅠb相似性仅为59.4%。结构分析结果表明,该基因可能具有几丁质酶/溶菌酶特点,且在过敏反应中起重要作用。
The complete cDNA sequence of the chitinase gene from wild banana leaves,was obtained by reverse transcription polymerase chain reaction （RT-PCR） with rapid amplification of cDNA ends （RACE）.The sequence had been submitted to the GenBank database （the accession number was FJ858155）.The full-length cDNA sequence of the wild banana was 1 115 bp,which contained a 924 bp open reading frame （ORF）,14 bp 5＇ untranslated regions,177 bp 3＇ untranslated regions and 17 bp poly （A） tail.The gene encoded a protein of 307 amino acids including a signal peptide,chitin-binding domain and glycoside-hydrolase domain.The estimated molecular weight and isoelectric point of the putative protein were 32 424.1 u and 6.28,respectively.The analysis showed that it was a transmembrane protein and secreted outside the cell.MuChiⅠ1 belongs to the acidic chitinase class Ib sub-family.Alignment of the chitinase class I in plantain （Musa paradisiacal L） showed that the sequence similarity was 59.4%.The structural analysis showed that the gene possessed the characteristic of chitinase and lysozyme,and it played an important role in the events of hypersensitive reaction （HR）.
出处
《热带作物学报》
CSCD
2010年第8期1244-1252,共9页
Chinese Journal of Tropical Crops
基金
福建省亚热带果树及特种经济作物种质资源共享平台资助项目部分内容(No.2008N2001)
国家科技支撑计划"闽台特色果树的离体试管保存
香蕉抗冻蛋白基因克隆及转基因"(No.2007BAD07B01)资助
关键词
野生香蕉
几丁质酶基因
基因克隆
序列分析
Wild banana （Musa spp., AB group）
Ehitinase gene
Gene cloning
Sequence analysis
