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引用本文:邓意龙,孙 雪,徐年军,杨 锐.蛋白核小球藻(Chlorella pyrenoidosa) Rubisco活化酶基因的克隆与表达分析.海洋与湖沼,2012,43(1):41-46.
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蛋白核小球藻(Chlorella pyrenoidosa) Rubisco活化酶基因的克隆与表达分析
邓意龙,孙 雪,徐年军,杨 锐
宁波大学应用海洋生物技术教育部重点实验室 宁波大学海洋学院
摘要:
采用RACE技术克隆了单细胞绿藻蛋白核小球藻820 Rubisco活化酶基因(rca)序列, 并用荧光定量PCR方法研究了rca和Rubisco小亚基基因(rbcS)的表达规律。 结果获得了1703bp的rcacDNA序列, 包括 66bp 的 5’非翻译区、1242bp 的开放阅读框和 395bp 的 3’非翻译区。序列比较和分析表明该蛋白核小球藻 rca 序列与其它绿藻的同源性高达 78%—85%; 偏好使用以 G/C/T 结尾的密码子; 推测的 RCA 蛋白等电点和分子量分别为 8.44 和 45.71kDa。 荧光定量 PCR 结果表明随光照时间增加rcarbcS 转录表达逐渐降低; 不同盐度处理下 rbcS 的 mRNA 量变化不大, 而 rca 在 37.5 盐度下表达量为 25 盐度的 2.69 倍; 1.0—3.0mmol/L 水杨酸处理 rcarbcS 表达均降低。
关键词:  蛋白核小球藻, Rubisco 活化酶基因, rbcS 基因, 转录表达
DOI:10.11693/hyhz201201006006
分类号:
基金项目:国家自然科学基金项目资助, 30700610 号; 浙江省公益性项目, 2010C33066 号
附件
THE CLONING AND EXPRESSION ANALYSIS OF RUBISCO ACTIVASE GENE IN THE UNICELLULAR GREEN ALGA CHLORELLA PYRENOIDOSA
DENG Yi-Long1,2, SUN Xue1,2, XU Nian-Jun1,2, YANG Rui1,2
1.Key Laboratory of Applied Marine Biotechnology, Minister of Education;2.School of Marine Sciences, Ningbo University
Abstract:
RACE technology was employed to clone the Rubisco activase gene (rca) of the unicellular green alga Chlorella pyrenoidosa 820, and the rca and rbcS (small subunit of Rubisco) expression profiles were investigated by real-time PCR. The obtained 1703bp rca cDNA sequence contained a 66bp 5’-untranslated region, a 1242bp open reading frame and a 395bp 3’-untranslated region. Sequence comparison showed that its homology with other green algae reached to 78%—85%. Sequence analysis showed that the rca gene preferred to use the codons ended with G or C or T, and the putative isoelectric point and molecular weight was 8.44 and 45.71kDa, respectively. Real-time PCR showed that rca and rbcS transcription quantities decreased as the light duration time increased. Under different salinity stress, rbcS mRNA quantities showed no significant variation, while rca mRNA quantities in 37.5 salinity reached to 2.69 fold of those in 25 salinity. The transcriptional expression of rca and rbcS all decreased after 1.0, 2.0 and 3.0mmol/L salicylic acid treatments.
Key words:  Chlorella pyrenoidosa, Rubisco activase gene, rbcS gene, Transcriptional expression
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