CAJ | 학술논문

利用已经公布的杉木444条EST序列和未公布的杉木基因组文库中1142条基因序列，进行引物开发效率的比较。去冗余后，利用MISA搜索SSR位点，分别得到109个和39个含有SSR的位点。杉木EST序列中SSR分布密度为964?58个/Mbp，基因组中平均每Mbp出现1037?24个SSR。在两个独立来源的数据库序列中，六核苷酸重复均为最多的重复类型，且AT-rich的重复类型占较大比例。 AGC/CTG是杉木EST序列和基因组库中最多的三碱基重复，通过Primer 3?0分别设计出SSR引物95对和37对。为考察设计引物在杉木不同种源（群体）中的有效性，取12个种源（个体）的优良个体，利用随机抽取的10个EST-SSR和8个gSSR （基因组SSR）进行引物筛选，结果表明：EST-SSR和gSSR各有4对引物在12个种源（个体）中表现出明显的多态性，多态率分别为40％和50％。8对多态性的SSR引物共扩增出25个多态性等位位点，平均每个引物产生3?125个多态性等位位点，平均有效的等位位点为2?3995，PIC平均值为0?5191；Hot平均为0?3074。其中gSSR标记在检测群体间存在较大的分化，4个gSSR比4个EST-SSR扩增出更多的等位位点数、平均等位位点数，以及更大的PIC值。
이용이경공포적삼목444조EST서렬화미공포적삼목기인조문고중1142조기인서렬，진행인물개발효솔적비교。거용여후，이용MISA수색SSR위점，분별득도109개화39개함유SSR적위점。삼목EST서렬중SSR분포밀도위964?58개/Mbp，기인조중평균매Mbp출현1037?24개SSR。재량개독립래원적수거고서렬중，륙핵감산중복균위최다적중복류형，차AT-rich적중복류형점교대비례。 AGC/CTG시삼목EST서렬화기인조고중최다적삼감기중복，통과Primer 3?0분별설계출SSR인물95대화37대。위고찰설계인물재삼목불동충원（군체）중적유효성，취12개충원（개체）적우량개체，이용수궤추취적10개EST-SSR화8개gSSR （기인조SSR）진행인물사선，결과표명：EST-SSR화gSSR각유4대인물재12개충원（개체）중표현출명현적다태성，다태솔분별위40％화50％。8대다태성적SSR인물공확증출25개다태성등위위점，평균매개인물산생3?125개다태성등위위점，평균유효적등위위점위2?3995，PIC평균치위0?5191；Hot평균위0?3074。기중gSSR표기재검측군체간존재교대적분화，4개gSSR비4개EST-SSR확증출경다적등위위점수、평균등위위점수，이급경대적PIC치。
In order to develop SSR primers of Cunninghamia lanceolata (Lamb.) Hook based on EST-SSR and genomic SSR, 292 non-repeated Chinese fir EST-sequences were assembled by removing low-quality and redundant fragments de-pend upon 444 ESTs sequences from NCBI Public database, and 143 genome-sequences were assembled by removing re-dundant fragments in 1 142 genome sequences. All those non-redundant sequences of Cunninghamia lanceolata (Lamb.) Hook was searched for mono-to hexa-nucleotide simple sequence repeats ( SSR or microsatellite) with software MISA, and the type,size and frequency of these SSRs were determined. Totally, There were 109 SSRs to be picked out among EST-sequences, the distribution density was 964.58 SSR/Mbp;and 39 SSRs were found in genome sequences, account-ing for the higher density of 1 037?24 SSR/Mbp. In both Genomic and EST sequence library, the Hexanucleotides re-peats, especially AT-rich repeat, was the most repeated type, and the AGC/CTG was the most trinucleotides repeats, The 37 and 95 pairs of SSR primers were designed based on EST library and Genomic by Primer 3.0, respectively. Ten EST-SSR primers and eight pairs of gSSR primers were selected randomly to perform PCR amplification with 12 individu-als. There were four pairs of primers showed polymorphism both in EST-SSR and gSSR, with polymorphic rate of 40%and 50%, respectively. Twenty-five polymorphic alleles were amplified by 8 SSR primer pairs, which showed an average 3?125 polymorphism alleles for each primer, the average effective alleles ( Na) 2.399 5, average PIC 0.519 1, average Hot 0?307 4. There were more polymorphism alleles from gSSR markers than EST-SSR among groups. More allelic loci were amplified with four pairs of primer from gSSR than the four pairs of primer from EST-SSR, and the former had high-er PIC value also.