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通过Southern 杂交等分析, 分离和克隆了龙眼叶绿体DNAL145 片段, 其中含大部分rbcL 基因。通过酶切分析制物理图谱, 结合亚克隆和PCR扩增, 测序分析了龙眼rbcL基因全序列1 836 bp , 其中基因编码区为1 428 bp, 编码475 个氨基酸和一个终止密码子, rbcL基因的5’上游区长284 bp , 在基因上游区- 191 ~- 220 bp 范围内有推测的类似原核生物的启动子序列: - 10 区的TACAAT, - 35 区的TTGCGC。在距离第一个转译起始密码子ATG 上游-6~- 10 bp 处, 有核糖体结合位点GGAGG, 类似于原核生物中的SD 序列。5’前导区185 bp。基因的3’下游区长124 bp, 在距离转译终止密码子TAA 下游30 ~100 bp 处, 有3 个转录茎环终止结构, 推测为终止子序列。将龙眼的rbcL 基因与烟草、水稻、葡萄、拟南芥菜等多种植物的相应序列进行了植物分子系统学的初步探讨。
By Southern hybridization analysis, the longan chloroplast DNAL145 fragment was isolated and cloned, which contains most of the rbcL gene. The full-length rbcL gene was analyzed by restriction enzyme digestion and PCR amplification. The full-length rbcL gene was 1 836 bp in length, including a 1 428 bp coding region of 475 amino acids and a stop codon. The rbcL gene The 5 ’upstream region is 284 bp in length. There are putative prokaryotic promoter sequences in the region of 191 - 220 bp upstream of the gene: TACAAT in region 10 and TTGCGC in region 35. At -6 to -10 bp upstream of the first translation initiation codon ATG, there is a ribosomal binding site GGAGG, similar to the SD sequence in prokaryotes. 5 ’leader 185 bp. The 3 ’downstream region of the gene is 124 bp in length. Three transcriptional stem loop termination structures are located 30 to 100 bp downstream from the translational stop codon TAA, suggesting a terminator sequence. The rbcL gene of longan and tobacco, rice, grape, Arabidopsis and other plants of the corresponding sequence of plant molecular phylogeny preliminary study.