甜橙β-胡萝卜素羟化酶cDNA克隆及其瞬间反义表达.pdfVIP

农业生物技术学报 Journal of Agricultural Biotechnology 2003,11 6冤院 593~597 ·研究论文· 甜橙 - 胡萝 卜素羟化酶cDNA 克隆及其瞬间反义表达* 昌杰 陈昆松 张上隆 秦巧平 (浙江大学园艺系，杭州310029) 摘要: 根据已报道的温州蜜柑( ) - 胡萝 卜素羟化酶( ) cDNA 序列(AF296158), 设计 1 对引物，以华盛顿 脐橙( )成熟果实cDNA 为模板，采用PCR 扩增出 1 条长为 1 057 bp 的cDNA 片段，将此片段克隆入pUCm-T 载体， 测序结果表明，所得序列与已报道序列同源99.62%, 表明所获得的 因是 因。将该 因反义插入到pBI121 中构建成 反义植物表达载体, 其正确性得到测序的鉴定。含有该植物表达载体的根癌农杆菌( )在脐橙白皮层 中诱导了 - 胡萝卜素积累。 关键词: 甜橙； - 胡萝 卜素羟化酶 因； 因克隆；反义植物表达载体; 瞬间表达 Cloning of -carotene Hydroxylase cDNA from Orange ( ) and Its Transient Antisense Expression Xu Changj ie Chen Kunsong Zhang Shanglong Qin Qiaoping (Department of Horticulture, Zheji ang University, Hangzhou 310029, China) A pair of non-degenerate primers, designed from known sequence of -carotene hydroxylase (GenBank AF296158) cDNA, was used to amplify same gene from orange （ ）with cDNA as template by polymerase chain reaction (PCR). As a result, an orange -carotene hydroxylase cDNA was obtained and sequenced. The nucleotide sequence of this gene from orange was 99.62% identical to the gene from . An antisense expression vector was constructed for this gene, and it was found that transient antisense expression of the gene in orange albedo successfully induced accumulation of -carotene. orange; -carotene hydroxylase gene; cloning; antisense plant expression vector; transient expression