カイコにおけるEST化したcDNAクローンのRFLPによる連関検索  [in Japanese] Linkage analysis of EST cDNA clones by using RFLP in the silkworm, Bombyx mori  [in Japanese]

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Abstract

cDNAクローンのRFLPを用いた連関検索法確立のために, サザン法によるシステムの検討を行った。3日齢卵のcDNAライブラリーよりプローブを調製した。数個のクローンを用いた解析で, 系統内多型が無く解析に適していたp50と日02号の両系統を用いて, プローブのRFLPの検索と連関検索を行った。その結果, 57%のクローンでRFLPにより差異が認められた。クローンの塩基配列を解析したところ, 5つがリボソームタンパク質遺伝子と, 4つが熱ショックタンパク質様アミノ酸配列をコードする遺伝子と類似であった。さらに, サザン法で明確なバンドを示さなかったものは, Bml, BMClに類似していた。p50と日02号のF<sub>1</sub>雌と日02号雄の戻し交雑の15個体のDNAを用いて極めて効率よく連関検索を行えることが判った。そこでこの方法をSLA法 (scanning linkage analysis method) と名付けた。連関検索の結果29クローンは, 9の連関群と8の独立したクローンとなった。

We started the linkage analysis of cDNA clones by using their restriction fragment length polymorphism (RFLP) and examined which strains should be crossed with, how to prepare the individual DNAs, probes, and so on. From the randomly picked-up plaques of 3 day-old embryonic cDNA library, EST-cDNA-clones of which insert size were longer than 500 by were subcloned into plasmid. The DIG ribo-probes were prepared by <i>in vitro</i> transcription and used for the Southern blot analysis. Some clones characterized already were used for screening the strains and the results indicated that p50 and J02 were isogenic. So we decided to use these strains and their cDNA-clones were analyzed. 57% of the clones were found to be useful for RFLP analyses. Sequence data for identification of clones gave new information, firstly, there were many unknowns and five ribosomal-protein-genes and four heat-shock-protein-genes, secondly, most of the clones that gave smear bands had repetitious sequences like Bm1 and Bm2. Linkage analyses of these clones were done by using the segregants of (p50×J02) female and J02 male. Every clone showing heterozygous or homozygous bands could be easily identified that they are linked or not. So this method was named scanning linkage analysis (SLA). Till now 29 cDNA clones were divided into 9 linkage groups and 8 independent clones out of 28 chromosomes.

Journal

  • The Journal of Sericultural Science of Japan

    The Journal of Sericultural Science of Japan 70(3), 135-143, 2001-12-27

    The Japanese Society of Sericultural Science

References:  41

Cited by:  7

Codes

  • NII Article ID (NAID)
    10010268133
  • NII NACSIS-CAT ID (NCID)
    AN00190300
  • Text Lang
    JPN
  • Article Type
    Journal Article
  • ISSN
    00372455
  • NDL Article ID
    6038552
  • NDL Source Classification
    ZR7(科学技術--農林水産--農産)
  • NDL Call No.
    Z18-343
  • Data Source
    CJP  CJPref  NDL  J-STAGE  JASI 
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