Knorr C, Cheng H H, Dodgson J B
Department of Microbiology, Michigan State University, East Lansing 48824, USA.
Anim Genet. 1999 Feb;30(1):28-35. doi: 10.1046/j.1365-2052.1999.00411.x.
The amplified fragment length polymorphism (AFLP) technique has been used to enhance marker density in the East Lansing reference chicken genome map, using a backcross family derived from a Red Jungle Fowl by White Leghorn mating with White Leghorn as the recurrent parent. To date, 204 AFLP markers have been added, expanding overall map coverage by about 25%. To the limits of our resolution, AFLP markers are distributed relatively evenly across the EL reference map. AFLP are about 60% as frequent in a cross within White Leghorns (line 7(2) x 6(3)) in comparison to the more divergent reference map population. Based on apparent identity of size, about 40% of the 7(2) x 6(3) cross AFLP fragments were also polymorphic in the reference map cross. Primer pairs in which one primer contains 3' extensions of three selective nucleotides and the other has two selective nucleotides successfully generated AFLP from chicken DNA, but such pairs appeared to amplify only a subset of those fragments to which they have an exact sequence match. Three different restriction enzymes with 4 bp recognition sites (TaqI, HinP1I and MspI) were found to work well with EcoRI as the rarer of the two AFLP restriction enzymes used, with HinP1I being the most effective of the three. AFLP markers are likely to provide an economical method with which to enhance framework linkage maps of chicken and probably other avian genomes.
扩增片段长度多态性(AFLP)技术已被用于提高东兰辛参考鸡基因组图谱的标记密度,该图谱使用了一个回交家系,该家系源自红原鸡与白来航鸡杂交,以白来航鸡作为轮回亲本。迄今为止,已添加了204个AFLP标记,使总图覆盖范围扩大了约25%。就我们的分辨率而言,AFLP标记在EL参考图谱上分布相对均匀。与差异较大的参考图谱群体相比,白来航鸡品系(7(2)×6(3))内杂交中的AFLP频率约为其60%。基于大小的明显一致性,7(2)×6(3)杂交中的AFLP片段约40%在参考图谱杂交中也是多态的。一对引物中一个引物含有三个选择性核苷酸的3'端延伸,另一个含有两个选择性核苷酸,成功地从鸡DNA中产生了AFLP,但这样的引物对似乎只扩增了与其有精确序列匹配的那些片段的一个子集。发现三种具有4 bp识别位点的不同限制酶(TaqI、HinP1I和MspI)与EcoRI配合良好,EcoRI是所用的两种AFLP限制酶中较稀有的一种,其中HinP1I是三种中最有效的。AFLP标记可能提供一种经济的方法来增强鸡以及可能其他鸟类基因组的框架连锁图谱。