摘要
目的:为获得凡纳滨对虾的热休克蛋白70(Hsp70)基因并分析其基因序列.方法:根据GenBank中斑节对虾(Penaeusmonodon)Hsp70基因的cDNA序列,设计引物,对经高盐法提取的凡纳滨对虾(Litopenaeusvannamei)基因组DNA,采用优化的降落PCR(TouchDownPCR)程序,扩增凡纳滨对虾Hsp70基因的全长序列.结果:PCR扩增得到一条长1983bp的目的DNA片段,回收纯化该片段并测定其核酸序列.用DNAman软件分析发现,该核酸序列中不含内含子,编码区全长为1959bp;经BLASTn和BLASTx软件分析发现,该编码区核苷酸序列与斑节对虾、罗氏沼虾(Macrobrachiumrosenbergii)的Hsp70基因序列的相似性分别为97%和62 2%.根据核苷酸序列所推导出的Hsp70氨基酸序列,其与斑节对虾、罗氏沼虾的相似性分别为99 9%和92 6%.结论:成功地从凡纳滨对虾基因组DNA中直接扩增出Hsp70基因的全长编码区序列.
Aim： To amplify the Hsp70 gene of Litopenaeus vannamei and study the features of the sequence. Methods：According to the Hsp70 cDNA sequence of Penaeus monodon accessed by GenBank, designed a pair of primers and used the optimal Touch Down PCR program to amplify the complete Hsp70 gene from genomic DNA of Litopenaeus vanamei. The total genomic DNA was extracted via the protocol of high - salt extraction. Results： A fragment of DNA, which was 1983 bp in size, was amplified and then sequenced. The sequence, analyzed with software DNAman, has a 1959 bp open reading frame, encoding 652 amino acids, but no intmn. With BLASTn and BLASTx from GenBank, the open reading frame of the Hsp70 gene sequence of Litopenaeus vannamei has 97% identity with the Hsp70 cDNA sequence of Penaeus monodon, and 62.2% identity with Macrobrachium rosenbergii. Under the alignment of Hsp70 amino acids deduced from DNA sequences, there is 99.9% identity between Litopenaeus vannamei and Penaeus monodon, and 92.6% identity between Litopenaeus vannamei and Macrobrachium rosenbergii. Conclusion： Directly and successfully amplified a complete Hsp70 gene from genomie DNA of Litopenaeus vannamei.
出处
《暨南大学学报（自然科学与医学版）》
CAS
CSCD
北大核心
2005年第3期428-433,共6页
Journal of Jinan University(Natural Science & Medicine Edition)
