Mapping flagellar genes in Chlamydomonas using restriction fragment length polymorphisms.
Mapping flagellar genes in Chlamydomonas using restriction fragment length polymorphisms.
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DOI：
10.1093/genetics/120.1.109
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发表时间：
1988-09
期刊：
影响因子：
3.3
通讯作者：
L. Ranum;Michael D. Thompson;J. A. Schloss;P. Lefebvre;C. Silflow
中科院分区：
文献类型：
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作者：
L. Ranum;Michael D. Thompson;J. A. Schloss;P. Lefebvre;C. Silflow
为了将克隆的核DNA序列与衣藻属先前特征的突变相关联，并深入了解其核基因组的组织结构，我们已经开始使用限制性片段长度多态(RFLP)来定位分子标记。在19个连锁群中的9个连锁群上含有表型标记的莱茵衣藻CC-29菌株与干扰种史密斯衣藻杂交。对随机选择的22个四分体的每个成员的DNA进行了分析，以确定与放射性DNA探针杂交检测到的克隆基因相关的RFLP。目前的标记集允许检测到与新的分子标记的连锁超过现有遗传图谱的大约54%。本研究的重点是定位克隆的鞭毛基因和转录产物在羽化后积累的基因。12个不同的分子克隆被归入7个连锁群。α-1微管蛋白基因定位于连接组III，并连接到与pcf6-100同源的基因组序列，pcf6-100是一个cdna克隆，其相应的转录产物在羽化后积累。α-2微管蛋白基因定位于连锁群IV。两个β-微管蛋白基因相连，其中β-1基因比β-2基因距离着丝粒更远约12 cM。与73kD动力蛋白相对应的克隆映射到同一连接组的另一臂。与基因克隆pcf6-187相对应的基因与连锁群V上紧密连锁的PF-26和PF-1突变非常接近，其mRNA在羽化后积累。
To correlate cloned nuclear DNA sequences with previously characterized mutations in Chlamydomonas and, to gain insight into the organization of its nuclear genome, we have begun to map molecular markers using restriction fragment length polymorphisms (RFLPs). A Chlamydomonas reinhardtii strain (CC-29) containing phenotypic markers on nine of the 19 linkage groups was crossed to the interfertile species Chlamydomonas smithii. DNA from each member of 22 randomly selected tetrads was analyzed for the segregation of RFLPs associated with cloned genes detected by hybridization with radioactive DNA probes. The current set of markers allows the detection of linkage to new molecular markers over approximately 54% of the existing genetic map. This study focused on mapping cloned flagellar genes and genes whose transcripts accumulate after deflagellation. Twelve different molecular clones have been assigned to seven linkage groups. The alpha-1 tubulin gene maps to linkage group III and is linked to the genomic sequence homologous to pcf6-100, a cDNA clone whose corresponding transcript accumulates after deflagellation. The alpha-2 tubulin gene maps to linkage group IV. The two beta-tubulin genes are linked, with the beta-1 gene being approximately 12 cM more distal from the centromere than the beta-2 gene. A clone corresponding to a 73-kD dynein protein maps to the opposite arm of the same linkage group. The gene corresponding to the cDNA clone pcf6-187, whose mRNA accumulates after deflagellation, maps very close to the tightly linked pf-26 and pf-1 mutations on linkage group V.