Isolation, expression and phylogenetic inheritance of an acetolactate synthase gene from Brassica napus
Isolation, expression and phylogenetic inheritance of an acetolactate synthase gene from Brassica napus
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甘蓝型油菜乙酰乳酸合酶基因的分离、表达及系统发育遗传
DOI：
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发表时间：
1989
期刊：
影响因子：
--
通讯作者：
M. Moloney
中科院分区：
文献类型：
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作者：
P. Wiersma;M. Schmiemann;J. Condie;W. Crosby;M. Moloney
从甘蓝型油菜中分离并鉴定了一个乙酰乳酸合成酶基因。这个B。napus乙酰乳酸合成酶基因编码637个氨基酸的多肽序列，与拟南芥的相应基因产物同源性为85%。最近与除草剂抗性/敏感性相关的肽结构域在两个序列之间是保守的。从Southern分析中，我们得出结论，该基因分离是一个多基因乙酰乳酸合成酶基因家族，包括四个或五个成员之一。Southern分析显示，跨越该基因的假定转运肽序列的探针与B中的独特序列杂交。油菜基因组这种独特的探针用于分析来自B的DNA。campestris和B. oleracea，推测是B的祖先。油菜。在限制性片段长度多态性的基础上，我们得出结论，B。napus基因起源于B。油菜在各种B中分析总乙酰乳酸脱氢酶同源转录物。油菜组织，并表现出优先积累在快速增长的材料。基因组克隆在体外在密码子173处突变以用丝氨酸替换脯氨酸残基。使用农杆菌载体将其重新引入植物中，产生除草剂抗性表型，这是预测的基因产物的特征。
SummaryAn acetolactate synthase gene was isolated and characterized from Brassica napus. This B. napus acetolactate synthase gene encodes a deduced polypeptide sequence of 637 amino acids which is 85% homologous to the corresponding proposed gene product from Arabidopsis thaliana. Peptide domains recently associated with herbicide resistance/sensitivity are conserved between the two sequences. From Southern analysis we conclude that the gene isolated is one member of a multigene acetolactate synthase gene family comprising four or five members. A probe spanning the presumptive transit peptide sequence of this gene was shown by Southern analysis to hybridize to a unique sequence in the B. napus genome. This unique probe was used to analyse DNA from B. campestris and B. oleracea, the presumed progenitors of B. napus. On the basis of restriction fragment length polymorphism, we conclude that the B. napus gene isolated here originated in B. campestris. Total acetolactate synthase-homologous transcripts were analysed in a variety of B. napus tissues, and showed preferential accumulation in rapidly growing material. The genomic clone was mutated in vitro at codon 173 to replace a proline residue with serine. This was re-introduced into plants, using Agrobacterium vectors, producing a herbicide-resistant phenotype which is characteristic of the predicted gene product.
“一种将 DNA 限制性内切酶片段放射性标记至高比活性的技术”。
影响因子：
2.9
作者：
Feinberg,AP;Vogelstein,B
通讯作者：
Vogelstein,B