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发明公开
CN103642822A 克隆苹果BKI1基因和苹果BKI1基因编码区序列的方法 失效 - 权利终止

克隆苹果BKI1基因和苹果BKI1基因编码区序列的方法
摘要：
本发明公开了克隆苹果BKI1基因和苹果BKI1基因编码区序列的方法。苹果BKI1基因的核苷酸序列如SEQIDNO:1所示，编码区序列如SEQIDNO:2所示，编码区序列编码氨基酸序列如SEQIDNO:3所示。苹果BKI1基因及其基因编码区序列的克隆方法，包括如下步骤：1）提取苹果叶片中的总DNA；2）提取苹果叶片中的总RNA反转录成cDNA；3）通过设计的特定引物对苹果的总DNA及cDNA分别进行PCR扩增反应；4）PCR产物回收和纯化；5）克隆，测序。本发明简单快捷的获取了苹果BKI1基因及其完整编码区序列，填补了该基因在苹果基因组注释上的空白，为研究苹果的矮化特性提供理论参考。
摘要（英）：

The invention discloses a method of cloning an apple BKI1 gene and an apple BKI1 gene coding region sequence. A nucleotide sequence of the apple BKI1 gene is shown as SEQIDNO:1, the coding region sequence is shown as SEQIDNO:2, and a coding amino acid sequence of the coding region sequence is shown as SEQIDNO:3. The method of cloning the apple BKI1 gene and the apple BKI1 gene coding region sequence comprises the steps: 1) extracting total DNA (Deoxyribonucleic Acid) in apple leaves; 2) extracting total RNA (Ribonucleic Acid) in the apple leaves, and reversely transcribing the total RNA into cDNA; 3) respectively carrying out PCR (Polymerase Chain Reaction) amplified reaction on the total DNA and cDNA of an apple by virtue of a designed specific primer; 4) recovering and purifying a PCR product; and 5) carrying out cloning and sequencing. With the adoption of the method, the apple BKI1 gene and the integral coding region sequence thereof are simply and rapidly obtained, the blank of the gene in annotation of an apple genome is filled, and theory reference is provided for studying the dwarfing characteristic of the apple.

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