水稻OsPIN1b基因GFP融合表达载体构建及转化水稻的研究
水稻生长素输出载体OsPIN1a可能参与调控水稻根负向光性反应。为了进一步研究其他OsPIN基因与水稻根负向光性的关系,以GenBank数据库报道的OsPIN1b核苷酸序列为基础,设计特异引物以RTPCR从水稻cDNA中扩增得到OsPIN1b基因。测序结果表明获得完整的OsPIN1b基因ORF序列;生物信息学分析表明,OsPIN1b基因序列全长1 665 bp,与已报道的序列相吻合,其GC含量为64.08%,编码554个氨基酸。进一步构建了该基因的GFP融合表达载体pCAMBIA-1301-OsPIN1b∷GFP并转化水稻中花11,分子检测和GUS染色表明外源片段已经成功整合到水稻基因组,为研...
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| Published in | 广东农业科学 Vol. 42; no. 24; pp. 146 - 150 |
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| Main Author | |
| Format | Journal Article |
| Language | Chinese |
| Published |
河南科技大学农学院,河南洛阳,471003%绍兴文理学院生命科学学院,浙江绍兴,312000
2015
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| Subjects | |
| Online Access | Get full text |
| ISSN | 1004-874X |
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| Summary: | 水稻生长素输出载体OsPIN1a可能参与调控水稻根负向光性反应。为了进一步研究其他OsPIN基因与水稻根负向光性的关系,以GenBank数据库报道的OsPIN1b核苷酸序列为基础,设计特异引物以RTPCR从水稻cDNA中扩增得到OsPIN1b基因。测序结果表明获得完整的OsPIN1b基因ORF序列;生物信息学分析表明,OsPIN1b基因序列全长1 665 bp,与已报道的序列相吻合,其GC含量为64.08%,编码554个氨基酸。进一步构建了该基因的GFP融合表达载体pCAMBIA-1301-OsPIN1b∷GFP并转化水稻中花11,分子检测和GUS染色表明外源片段已经成功整合到水稻基因组,为研究OsPIN1b在水稻根负向光性中的作用奠定基础。 |
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| Bibliography: | 44-1267/S rice（Oryza sativa）; OsPIN1b; negative phototropism XU Hua-wei;WANG Hai;MO Yi-wei（ 1. College of Agriculture, Henan University of Science and Techen.ology, Luoyang 471003, China; 2. School of Life Sciences, Shaoxing University, Shaoxing 312000, China ） Earlier research results showed that rice auxin efflux protein OsPIN1 a probably participated in regulating negative phototropism of rice roots. To explore the relationship between other OsPIN genes and negative phototropism of rice roots,gene-specific primers were used to amplify Os PIN1 b gene by RT-PCR based on the OsPIN1 b sequence in Gen Bank. Sequencing result showed that the full ORF was obtained（1 665 bp）. Bioinformation analysis showed that the sequence was 100% identity with the reported sequence,the GC content of OsPIN1 b ORF was 64.08% and OsPIN1 b encoded 554 amino acid residues. The OsPIN1b-green fluorescent protein（gfp）fusion expression vector p CAMBIA-1301-Os PIN1b∷GFP was constructed and transformed into Japonicarice Zhonghua11 by Agrobac |
| ISSN: | 1004-874X |