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湿地松及其他裸子植物中作为大基因组组成部分的类Ty1-copia序列的基因组和物理组织

The genomic and physical organization of Ty1-copia-like sequences as a component of large genomes in Pinus elliottii var. elliottii and other gymnosperms.

作者信息

Kamm A, Doudrick R L, Heslop-Harrison J S, Schmidt T

机构信息

Department of Cell Biology, John Innes Institute, Norwich United Kingdom.

出版信息

Proc Natl Acad Sci U S A. 1996 Apr 2;93(7):2708-13. doi: 10.1073/pnas.93.7.2708.

Abstract

A DNA sequence, TPE1, representing the internal domain of a Ty1-copia retroelement, was isolated from genomic DNA of Pinus elliottii Engelm. var. elliottii (slash pine). Genomic Southern analysis showed that this sequence, carrying partial reverse transcriptase and integrase gene sequences, is highly amplified within the genome of slash pine and part of a dispersed element >4.8 kbp. Fluorescent in situ hybridization to metaphase chromosomes shows that the element is relatively uniformly dispersed over all 12 chromosome pairs and is highly abundant in the genome. It is largely excluded from centromeric regions and intercalary chromosomal sites representing the 18S-5.8S-25S rRNA genes. Southern hybridization with specific DNA probes for the reverse transcriptase gene shows that TPE1 represents a large subgroup of heterogeneous Ty1-copia retrotransposons in Pinus species. Because no TPE1 transcription could be detected, it is most likely an inactive element--at least in needle tissue. Further evidence for inactivity was found in recombinant reverse transcriptase and integrase sequences. The distribution of TPE1 within different gymnosperms that contain Ty1-copia group retrotransposons, as shown by a PCR assay, was investigated by Southern hybridization. The TPE1 family is highly amplified and conserved in all Pinus species analyzed, showing a similar genomic organization in the three- and five-needle pine species investigated. It is also present in spruce, bald cypress (swamp cypress), and in gingko but in fewer copies and a different genomic organization.

摘要

从湿地松(Pinus elliottii Engelm. var. elliottii)的基因组DNA中分离出一个代表Ty1 - copia逆转录转座子内部结构域的DNA序列TPE1。基因组Southern分析表明,该序列携带部分逆转录酶和整合酶基因序列,在湿地松基因组中高度扩增,是一个大于4.8 kbp的分散元件的一部分。对中期染色体进行荧光原位杂交显示,该元件相对均匀地分布在所有12对染色体上,且在基因组中含量丰富。它在很大程度上被排除在着丝粒区域和代表18S - 5.8S - 25S rRNA基因的染色体中间位点之外。用逆转录酶基因的特异性DNA探针进行Southern杂交表明,TPE1代表了松属物种中异质Ty1 - copia逆转录转座子的一个大亚组。由于未检测到TPE1转录,它很可能是一个无活性元件——至少在针叶组织中是这样。在重组逆转录酶和整合酶序列中发现了无活性的进一步证据。通过Southern杂交研究了PCR分析所示的TPE1在含有Ty1 - copia类逆转录转座子的不同裸子植物中的分布。TPE1家族在所有分析的松属物种中高度扩增且保守,在所研究的三针松和五针松物种中显示出相似的基因组结构。它也存在于云杉、落羽杉(沼柏)和银杏中,但拷贝数较少且基因组结构不同。

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