Ezaz M Tariq, Harvey Simon C, Boonphakdee Chuta, Teale Alan J, McAndrew Brendan J, Penman David J
Institute of Aquaculture, University of Stirling, Stirling FK9 4LA, Scotland, UK.
Mar Biotechnol (NY). 2004 Sep-Oct;6(5):435-45. doi: 10.1007/s10126-004-3004-6. Epub 2004 Aug 24.
Gynogenetically produced XX and YY Nile tilapia (Oreochromis niloticus) and diploid control groups were screened for amplified fragment length polymorphisms (AFLPs) to search for sex-linked or sex-specific markers. Family-level bulked segregant analysis (XX and YY gynogenetic family pools) and individual screening (XX and YY gynogenetics and XX and XY control individuals) identified 3 Y-linked (OniY425, OniY382, OniY227) and one X-linked (OniX420) AFLP markers. OniX420 and OniY425 were shown to be allelic. Single locus polymerase chain reaction assays were developed for these markers. Tight linkage was demonstrated between the AFLP markers and the sex locus within the source families. However, these markers failed to consistently identify sex in unrelated individuals, indicating recombination between the markers and the sex-determining loci. O. niloticus bacterial artificial chromosome clones, containing the AFLP markers, hybridized to the long arm of chromosome 1. This confirmed previous evidence, based on meiotic chromosome pairing and fluorescence in situ hybridization probes obtained through chromosome microdissection, that chromosome pair 1 is the sex chromosomes.
对雌核发育产生的XX和YY尼罗罗非鱼（Oreochromis niloticus）以及二倍体对照组进行扩增片段长度多态性（AFLP）筛选，以寻找性别连锁或性别特异性标记。通过家系水平的混合分离分析（XX和YY雌核发育家系池）和个体筛选（XX和YY雌核发育个体以及XX和XY对照个体），鉴定出3个Y连锁（OniY425、OniY382、OniY227）和1个X连锁（OniX420）AFLP标记。结果表明OniX420和OniY425是等位基因。针对这些标记开发了单基因座聚合酶链反应检测方法。在源家系中证实了AFLP标记与性别位点之间存在紧密连锁。然而，这些标记未能在无关个体中始终如一地鉴定性别，这表明标记与性别决定位点之间发生了重组。含有AFLP标记的尼罗罗非鱼细菌人工染色体克隆与第1号染色体的长臂杂交。这证实了先前基于减数分裂染色体配对和通过染色体显微切割获得的荧光原位杂交探针得出的证据，即第1号染色体对是性染色体。
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