利用mRNA差别显示技术分离盐胁迫下红树植物白骨壤耐盐相关cDNA  

Extraction of the Salt-tolerant cDNA in Mangrove Avicennia marina by mRNA Differential Display

作　　者：周涵韬[1] 林鹏[1] 

机构地区：[1]厦门大学生命科学学院,厦门361005

出　　处：《生物工程学报》2002年第1期51-54,共4页Chinese Journal of Biotechnology

基　　金：国家教育部高等学校博士点基金资助 (No.19990 38410 )~~

摘　　要：从福建龙海红树林自然保护区采集白骨壤的隐胎生果实 ,在实验室分别置于 0‰盐度海水和 5 0‰盐度海水进行沙培。分别取叶片 ,提取纯化RNA。通过锚定引物OligodT1 2 GC反转录和 8个 10核苷酸随机引物进行PCR扩增 ,经 8%非变性聚丙烯酰胺凝胶电泳后发现了 3个差异DNA片段只在高盐培养条件的白骨壤基因组中表达 ,而在无盐培养条件中没有出现。这 3个差异cDNA片段分别命名为csrg1(6 0 0bp)、csrg2 (5 5 0bp)、csrg3(4 80bp)。 3个差异cDNA片段的RNA杂交结果显示 ,只有csrg1片段存在明显差异 高盐中有杂交斑点 ,无盐中无杂交斑点 ;而其余 2个片段在高盐和无盐条件下都没有杂交斑点出现。从而表明csrg1就是耐盐相关cDNA。进一步将csrg1片段克隆 ,并进行DNA序列分析。全序列在GenBank中查询后 ,未发现相关同源片段。耐盐相关cDNA片段的获得 ,将为分离全长耐盐基因 。RNAs of the leaves in \%Avicennia marina,\% which cultured in 50‰ and 0‰ salinity condition respectively, were isolated for mRNA differential display analysis. Screened by OligodT 12 GC and eight 10-oligonucleotide arbitrary primers,differential cDNA fragments-csrg1(600bp)\,csrg2(550bp)\,csrg3(480bp),only appeared in the leaves of \%Avicennia marina\% cultured in 50‰ salinity condition.After detected by RNA dot hybridization, only csrg1 appeared the difference between the RNAs of the leaves in \%Avicennia marina\% cultured in 50‰ and 0‰ salinity condition respectively, and csrg1 was confirmed as the salt-tolerant cDNA.csrg1 was cloned and sequenced.After searching in Genbank,there were no similar sequences reported.

关 键 词：MRNA差别显示技术 分离 盐胁迫 红树植物 白骨壤 耐盐相关cDNA 耐盐性 

分 类 号：Q945.78[生物学—植物学]