Houben A, Kynast R G, Heim U, Hermann H, Jones R N, Forster J W
Institut fur Pflanzengenetik und Kulturpflanzenforschung, D-06466 Gatersleben, Germany.
Chromosoma. 1996 Aug;105(2):97-103. doi: 10.1007/BF02509519.
The terminal heterochromatic segments of the long arms of 20 rye B-chromosomes were isolated by means of laser microdissection technology. Also the remaining portions of the long arms, along with the short arms of the same chromosomes were isolated. Each sample was used for degenerate oligonucleotide primer-polymerase chain reaction (DOP-PCR) amplification reactions. The resulting products were used as probes for chromosome in situ hybridisation experiments, and in Southern hybridisation to digests of 0B and +B DNA. Competition hybridisation of these probes with 0B DNA allowed the detection of B-specific sequences. The terminal heterochromatin of the rye B-chromosome contains both B-specific sequences and sequences also present on the A-chromosomes of rye. The B-specific D1100 family is the major repeat species located in the terminal heterochromatin. Primers designed to the cloned sequence (E1100) were used to search for related low copy sequences in 0B DNA. The sequences of the PCR products revealed no similarities to that of the clone E1100 except for the primer sequences. The possible origin of this sequence is discussed in the context of models for the evolution of the rye B-chromosome.
通过激光显微切割技术分离出20条黑麦B染色体长臂的末端异染色质区段。同样，这些染色体长臂的其余部分以及短臂也被分离出来。每个样本用于简并寡核苷酸引物聚合酶链反应（DOP-PCR）扩增反应。所得产物用作染色体原位杂交实验的探针，并用于对0B和+B DNA酶切产物的Southern杂交。这些探针与0B DNA的竞争杂交使得能够检测到B特异性序列。黑麦B染色体的末端异染色质既包含B特异性序列，也包含黑麦A染色体上也存在的序列。B特异性D1100家族是位于末端异染色质中的主要重复序列类型。针对克隆序列（E1100）设计的引物用于在0B DNA中寻找相关的低拷贝序列。PCR产物的序列除引物序列外与克隆E1100的序列没有相似性。在黑麦B染色体进化模型的背景下讨论了该序列的可能起源。
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