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題 名 | 豬基因組遺傳標誌--微衛星之選殖與分析=Cloning and Analysis of the Genetic Marker of Repetitive Sequence: Microsatellite DNA in Porcine Genome |
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作 者 | 鄭益謙; 劉名允; 陳啟銘; | 書刊名 | 中華民國獸醫學會雜誌 |
卷 期 | 25:3 1999.09[民88.09] |
頁 次 | 頁 218-225 |
分類號 | 437.22 |
關鍵詞 | 豬基因庫; 遺傳標誌; 微衛星; 聚合酶鏈反應; 多型性; Pig genome; Genetic marker; Microsatellite; Polymerase chain reaction; Polymorphism; |
語 文 | 中文(Chinese) |
中文摘要 | 我們以質體建立豬染色體片段之隨機基因庫(random library),進行豬基因庫微衛星之選殖。從基因庫篩選可能含有(CA)序列重複的50個選殖株進行定序,得到24株具(CA)序列重複的片段選殖株,其中兩株另含(A)重複序列,另有一選殖株不含(CA)序列重複,而為含48個(T)重複之片段。選殖株中豬染色體各片段經定序後發現(CA)序列重複次數為10-26次。針對其中9個含(CA)及唯一一個(T)重複序列染色體選殖片段序列設計引子,並設定聚合�t鏈反應最佳條件。此10對引子(PRT-1∼PRT-10)聚合�t鏈反應之產物長度為154-258bp,以之與64頭豬(藍瑞斯、杜洛克、約克夏、盤克夏、小耳種豬、桃園豬和山豬)之gDNA模版行聚合�t鏈反應,檢測重複序列之多型性,發現各對引子產生之基因型個數為2-10個,共產生66個基因型;10個重複序列之多型性指數(PIC, olymorphism information content)除PRT-1為0.09外,其餘均在0.5以上(0.55-0.84),在本試驗中並未發現(CA)序列重複次數之多寡與基因型個數有相關性。再以一個藍瑞斯家族進行基因型遺傳檢定,這10個選殖片段基因型均符合孟德爾遺傳律,因此我們認為內含微衛星、PIC大於0.5的9個選殖片段均適合作為動物經濟性狀篩選或親緣鑑定等的遺傳標誌。 |
英文摘要 | We report here the construction of a porcine random genomic library by plasmids and sequencing of 50 clones possibly containing (CA) repeats. Twenty-four clones contained (CA) repeats among which two clones with additional A nucleotides were obtained. However, one clone with 48 additional T nucleotides within the sequence was also observed. The range of (CA) repeats of clones with porcine genomic fragments was from 10 to 26. The primers flanking these 9 clones with (CA) repeats and the clone with additional T nucleotides were selected for test. Tests for optimal polymerase chain reaction (PCR) amplification of the repeats were conducted. These PCR products of the 10 clones amplified were in the range of 154 to 258 base pairs. To test the polymorphism of these 10 clones, sixty-four gDNA samples including Landrace, Yorkshire, Duroc, Berkshire, Native small-ear pig, Taoyuan pig, and Wild boar were amplified by standard PCR with pairs of primers to check the polymorphism of repetitive sequences, so called microsatellites. The genotype number of the PCR products amplified by the primer each ranged from two to ten. Totally, there were sixty-six genotypes obtained from these ten primer pairs. Except for the (CA) repeats named PRT-1 with a PIC polymorphism information content) of 0.09, the PIC of the other microsatellites were all above 0.5 (0.55-0.84). There was no correlation between the (CA) repeat number and the genotype number in our experiment. To prove that the (CA) repeat of the offspring was inherited from their parents, these ten primers were tested on one Landrace family. The results of the pedigree study indicated that the inheritance followed the Mendelian principles. Therefore, these nine clones with PIC values above 0.5 can be applied for the purposes such as the screening of the traits of commercial interests or identifying the relationships among animals. |
本系統中英文摘要資訊取自各篇刊載內容。