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质体小环的复制涉及滚环中间体。

The replication of plastid minicircles involves rolling circle intermediates.

作者信息

Leung Siu Kai, Wong Joseph T Y

机构信息

Department of Biology, Hong Kong University of Science and Technology, Kowloon, Hong Kong SAR, PR China.

出版信息

Nucleic Acids Res. 2009 Apr;37(6):1991-2002. doi: 10.1093/nar/gkp063. Epub 2009 Feb 10.

DOI:10.1093/nar/gkp063
PMID:19208639
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC2665238/
Abstract

Plastid genomes of peridinin-containing dinoflagellates are unique in that its genes are found on multiple circular DNA molecules known as 'minicircles' of approximately 2-3 kb in size, carrying from one to three genes. The non-coding regions (NCRs) of these minicircles share a conserved core region (250-500 bp) that are AT-rich and have several inverted or direct repeats. Southern blot analysis using an NCR probe, after resolving a dinoflagellate whole DNA extract in pulsed-field gel electrophoresis (PFGE), revealed additional positive bands (APBs) of 6-8 kb in size. APBs preferentially diminished from cells treated with the DNA-replication inhibitor aphidicolin, when compared with 2-3 kb minicircles, implicating they are not large minicircles. The APBs are also exonuclease III-sensitive, implicating the presence of linear DNA. These properties and the migration pattern of the APBs in a 2D-gel electrophoresis were in agreement with a rolling circle type of replication, rather than the bubble-forming type. Atomic force microscopy of 6-8 kb DNA separated by PFGE revealed DNA intermediates with rolling circle shapes. Accumulating data thus supports the involvement of rolling circle intermediates in the replication of the minicircles.

摘要

含多甲藻素的甲藻的质体基因组很独特，其基因存在于多个环状DNA分子上，这些分子被称为“微环”，大小约为2 - 3 kb，每个微环携带一到三个基因。这些微环的非编码区（NCRs）共享一个保守的核心区域（250 - 500 bp），该区域富含AT，并有几个反向或正向重复序列。在脉冲场凝胶电泳（PFGE）中分离甲藻全DNA提取物后，用NCR探针进行Southern印迹分析，结果显示有大小为6 - 8 kb的额外阳性条带（APBs）。与2 - 3 kb的微环相比，用DNA复制抑制剂阿非迪霉素处理的细胞中，APBs优先减少，这表明它们不是大的微环。APBs对外切核酸酶III也敏感，这表明存在线性DNA。这些特性以及APBs在二维凝胶电泳中的迁移模式与滚环型复制一致，而不是气泡形成型。通过PFGE分离的6 - 8 kb DNA的原子力显微镜观察显示出具有滚环形状的DNA中间体。因此，越来越多的数据支持滚环中间体参与微环的复制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7d39/2665238/ea23e944bca9/gkp063f8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7d39/2665238/556bd07204c6/gkp063f1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7d39/2665238/63b22fae80d6/gkp063f2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7d39/2665238/9dd7bf45a3b2/gkp063f3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7d39/2665238/20e35d15539c/gkp063f4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7d39/2665238/d633c94e9ba4/gkp063f5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7d39/2665238/f616fd93abdf/gkp063f6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7d39/2665238/58ac4e759750/gkp063f7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7d39/2665238/ea23e944bca9/gkp063f8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7d39/2665238/556bd07204c6/gkp063f1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7d39/2665238/63b22fae80d6/gkp063f2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7d39/2665238/9dd7bf45a3b2/gkp063f3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7d39/2665238/20e35d15539c/gkp063f4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7d39/2665238/d633c94e9ba4/gkp063f5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7d39/2665238/f616fd93abdf/gkp063f6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7d39/2665238/58ac4e759750/gkp063f7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7d39/2665238/ea23e944bca9/gkp063f8.jpg

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