| 胡亚平，郭雁君，吉前华，蒋 惠，郭丽英，张小凤，周希琴.砂糖桔全长cDNA 文库的构建及病程
相关蛋白基因的克隆与分析[J].广东农业科学,2015,42(16):89-88 |
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砂糖桔全长cDNA 文库的构建及病程
相关蛋白基因的克隆与分析 |
| Construction of a full-length cDNA library of Shatangjuand cloning of pathogenesis-related protein gene |
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| DOI： |
| 中文关键词: 砂糖桔 cDNA 文库 病程相关蛋白 |
| 英文关键词: Citrus flamea Hort. ex Tseng. Shiyueju cDNA library pathogenesis-related protein |
| 基金项目:肇庆学院自然科学基金（201103）；肇庆
学院科研创新团队项目（201205） |
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| 摘要点击次数: 1648 |
| 全文下载次数: 774 |
| 中文摘要: |
| 为从分子水平上了解影响砂糖桔生理代谢、生长发育、病虫害抗性和品质的众多基因，以无籽
砂糖桔幼叶为材料，应用SMART 技术构建了cDNA 文库，检测文库发现其库容量达到1.36×106，重组率为
95%，插入片段绝大多数分布在0.6~2 kb 之间，有36% 的插入片段是全长cDNA。随机挑选224 个克隆进行测
序，获得了1 个病程相关蛋白基因的全长cDNA 序列。将基因序列提交GenBank，编号为KJ000019。基因的
CDS 长429 bp，5′UTR 为43 bp，3′UTR 为235 bp，编码长度为143 个氨基酸残基的蛋白。基因位于第8 染色
体上，有1 个长度为193 bp 的内含子，基因编码的蛋白质分子量为15.4 ku，等电点为6.96。该蛋白N 端含有
长度约为22 aa 的信号肽序列，指导蛋白分泌到细胞外；剩余部分是一个典型的Barwin 保守结构域，具有几丁
质酶的活性，在受到细菌、真菌侵染时可以分解部分真菌和细菌的细胞壁，发挥抗病作用。 |
| 英文摘要: |
| To better understand the molecular mechanisms underlying metabolism，development，disease
resistance and quality of citrus，we constructed a full-length cDNA library from Citrus Shatangju（seedless）by
using SMART™ technology. This high quality cDNA library featured as a storage capacity of 1.36 × 106 with more
than 95% recombinant，containing a wide range of insert size from 0.6-2 kb. 224 clones were randomly picked up
from the library and sequenced，36% of the clones were full length cDNAs. One of them，pathogenesis-related protein
gene was submitted to GenBank with accesses number KJ000019. Bioinformatics analysis found that the full-length of
pathogenesis-related protein cDNA was 707 bp，with a CDS of 459 bp，a 43 bp 5′UTR and a 235 bp 3′UTR，encoding
a protein with 143 amino acid residues，the corresponding genomic sequence contained one intron with 193 bp. The
molecular weight of the protein was 15.4 ku，with isoelectric point（pI）value of 6.96. The pathogenesis-related protein
was predicted to have a signal peptide with 22 amino acid residences，which guided the protein to secreted to the outside
of cell，and a Barwin domain，which responsed to fungi and bacteria with chitinase activity. The results indicated that，
construction of this high quality cDNA library would be valuable to clone candidate genes，enhancing our understanding
to the physiological metabolic processes of citrus，the obtained pathogenesis-related protein was valuable to diseaseresistant
engineering. |
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