【摘要】 【目的】家蚕(Bombyx moriL.)品种和纯度的鉴别,普遍依靠幼虫斑纹或茧形等形态鉴别方法,该方法易受环境影响,结果时效性和准确性差。【方法】筛选RAPD随机引物,稳定扩增出亲本品种的DNA特异性片段,转换成SCAR标记。【结果】利用微量DNA抽提法,从家蚕主要品种苏5、苏6及其F1孵化前日卵快速提取单粒卵基因组DNA,筛选出RAPD随机引物S42,稳定扩增出苏5和苏6两个亲本品种的DNA特异性片段R5和R6,克隆和测序确认其大小分别为255bp和343bp。Blastn分析显示,R5的nt7～192与家蚕的一个非长末端逆转录转座子(AB002270.1)的nt183～368片段碱基序列有高达91%的相似性,无缺口序列。R6的nt5～340与家蚕的一个WGS(BAAB01113163.1)的nt675～337有91%的一致性,其中存在13个碱基的缺口序列。将2个亲本的RAPD标记转换成SCAR标记,分别利用合成的S42-255-2(P5-2)和S42-343-2(P6-2)SCAR标记引物,能够准确、快速地鉴别所标记的苏5和苏6及其F1蚕品种。【结论】SCAR分子标记方法能够检测家蚕品种和纯度。更多还原

【Abstract】 【Objective】The rule method to identify silkworm (Bombyx mori) variety and purity with morphological characters of larval pattern or cocoon shape is easily affected by environment. Thus,the prescription and accuracy are low. 【Method】Select the random primers by random amplified polymorphic DNA (RAPD), amplify the specific bands of RAPD from two parental varieties, then transform the RAPD marks to sequence characterized amplified regions (SCAR) marks. 【Result】The genome DNA was extracted from single blue-stage-egg of mainly Bombyx varieties of SU5, SU6 and their F1with rapid method. Each of the specific bands R5 and R6 of RAPD were amplified stably from SU5 and SU6 varieties, with selected random primer of S42. The sequences of bands R5 and R6 were 255 bp and 343 bp respectively after being cloned and sequenced. Then analyze the sequences by courtesy of database on-line. The results showed the nt 7-192 of R5 and the nt 183-368 of the non-LTR retrotransposon of Bombyx mori L1Bm DNA (AB002270.1) have 91% consistency, with no gap sequence (Gaps). Accordingly, the nt 5-340 of R6 and nt 675-337 of one of whole genome shotgun sequences (BAAB01113163.1| Bombyx mori DNA, contig491593) have 91% consistency, which include 13 gaps. Further, transform the RAPD marks of two parental varieties to SCAR marks. Thus, SU5, SU6 and their F1 were identified accurately and quickly, with the synthetic specific primers of S42-255-2 (P5-2) and S42-343-2 (P6-2). 【Conclusion】Molecular mark of SCAR can be used for identifying the variety and purity of silkworm, Bombyx mori L.更多还原