中国龙虾微卫星标记的筛选及遗传多样性分析Screeningandgenetic.PDF

HEREDITAS (Beijing) ISSN 0253-9772 中国龙虾微卫星标记的筛选及遗传多样性分析∗ 刘楚吾，黎锦明，刘丽，郭昱嵩 广东海洋大学水产学院，南海水产经济动物增养殖广东普通高校重点实验室，湛江 524025 摘要：文章以M13 通用引物和重复序列(CT)15、(AT)15 引物，利用 PCR 法对中国龙虾（Panulirus stimpsoni Hoehuis ）部分基 因组DNA 文库进行筛选。共获得 78 个微卫星序列，分别分布于 55 个阳性重组克隆中，其中完美型（perfect ）共50 个，占 64%；非完美型 （imperfect ）3 个，占3.8% ；混合完美型 （compound perfect ）6 个，占7.7% ；混合非完美型 （compound imperfect ） 19 个，占 24.5% 。根据微卫星序列，设计并筛选出 15 对微卫星多态性引物，对中国龙虾的群体进行了遗传多样性分析。获 得 3～12 个等位基因，等位基因大小在 78～425 bp 之间，基本符合引物设计的理论长度。期望杂合度范围为 0.48～0.87，平 均值为 0.71，表明中国龙虾基因组微卫星具有较高的杂合度与遗传多样性。15 个微卫星位点的 PIC 值从 0.44 到 0.84，平均 值为 0.60，说明这些微卫星位点在中国龙虾基因组中包含丰富的遗传信息，合适用于中国龙虾的各种分子标记及遗传学分析 和应用。 关键词：中国龙虾；基因组文库；微卫星标记；遗传多样性；引物筛选 Screening and genetic diversity analysis of microsatellite markers in Chinese lobster （Panulirus stimpsoni ） LIU Chu-Wu, LI Jin-Ming, LIU Li, GUO Yu-Song Fisheries College, Key Labolatory of Aquaculture in South China Sea for Aquatic Economic Animal of Guangdong Higher Education Institutes,Guangdong Ocean University, Zhanjiang 524025, China Abstract: With the construction of a library of partial fractionated genomic DNA of Panulirus stimpsoni Hoehuis, the microsatellite sequences of P. stimpsoni were screened by PCR technique. Then, the genetic diversity was analyzed with the microsatellite markers. Seventy-eight microsatellite sequences in 55 positive recombinant clones were obtained by PCR technique with primers of M13+/- and (CT)15, and (AT)15. Among these microsatellite sequences, the numbers of perfect, imperfect, compound perfect, and compound imperfect sequences were 50 (64%), 3 (3.8%), 5 (7.7%), and 19 (24.5%), respectively. To analyze genomic DNA diversity of P. stimpsoni, 15 pairs of primers were designed from the microsatellite flanking sequences. In these microsatellite loci, the alleles numbers ranged from 3 to 12; and the