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大麦中的Nir1位点与亚硝酸还原酶脱辅基蛋白基因Nii紧密连锁。

The Nir1 locus in barley is tightly linked to the nitrite reductase apoprotein gene Nii.

作者信息

Ward M P, Abberton M T, Forde B G, Sherman A, Thomas W T, Wray J L

机构信息

Plant Sciences Laboratory, School of Biological and Medical Sciences, University of St. Andrews, Fife, UK.

出版信息

Mol Gen Genet. 1995 Jun 10;247(5):579-82. doi: 10.1007/BF00290349.

Abstract

pBNiR1, a cDNA clone encoding part of the barley nitrite reductase apoprotein, was isolated from a barley (cv. Maris Mink) leaf cDNA library using the 1.85 kb insert of the maize nitrite reductase cDNA clone pCIB808 as a heterologous probe. The cDNA insert of pBNiR1 is 503 bp in length. The nucleotide coding sequence could be aligned with the 3' end of other higher plant nitrite reductase apoprotein cDNA sequences but diverges in the 3' untranslated region. The whole-plant barley mutant STA3999, previously isolated from the cultivar Tweed, accumulates nitrite after nitrate treatment in the light, has very much lowered levels of nitrite reductase activity and lacks detectable nitrite reductase cross-reacting material due to a recessive mutation in a single nuclear gene which we have designated Nir1. STA3999 has the characteristics expected of a nitrite reductase apoprotein gene mutant. Here we have used pB-NiR1 in RFLP analysis to determine whether the mutation carried by STA3999 is linked to the nitrite reductase apoprotein gene locus Nii. An RFLP was identified between the wild-type barley cultivars Tweed (major hybridising band of 11.5 kb) and Golden Promise (major hybridising band of 7.5 kb) when DraI-digested DNA was probed with the insert from the partial barley nitrite reductase cDNA clone, pBNiR1. DraI-digested DNA from the mutant STA3999 also exhibited a major hybridising band of 11.5 kb after hybridisation with the insert from pBNiR1.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

pBNiR1是一个编码大麦亚硝酸还原酶脱辅基蛋白部分序列的cDNA克隆,它是利用玉米亚硝酸还原酶cDNA克隆pCIB808的1.85 kb插入片段作为异源探针,从大麦(品种Maris Mink)叶片cDNA文库中分离得到的。pBNiR1的cDNA插入片段长度为503 bp。其核苷酸编码序列可与其他高等植物亚硝酸还原酶脱辅基蛋白cDNA序列的3'端比对,但在3'非翻译区存在差异。之前从品种Tweed中分离出的全株大麦突变体STA3999,在光照下经硝酸盐处理后会积累亚硝酸盐,其亚硝酸还原酶活性水平大幅降低,并且由于一个单细胞核基因(我们命名为Nir1)中的隐性突变,缺乏可检测到的亚硝酸还原酶交叉反应物质。STA3999具有亚硝酸还原酶脱辅基蛋白基因突变体所预期的特征。在此,我们利用pB-NiR1进行限制性片段长度多态性(RFLP)分析,以确定STA3999携带的突变是否与亚硝酸还原酶脱辅基蛋白基因位点Nii连锁。当用部分大麦亚硝酸还原酶cDNA克隆pBNiR1的插入片段探测经DraI酶切的DNA时,在野生型大麦品种Tweed(主要杂交带为11.5 kb)和Golden Promise(主要杂交带为7.5 kb)之间鉴定出了一个RFLP。用pBNiR1的插入片段杂交后,突变体STA3999经DraI酶切的DNA也显示出一条11.5 kb的主要杂交带。(摘要截短至250字)

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