Ruf M, Kössel H
Institut für Biologie III, Universität Freiburg, FRG.
Nucleic Acids Res. 1988 Jul 11;16(13):5741-54. doi: 10.1093/nar/16.13.5741.
The rpoA gene coding for the alpha-subunit of DNA-dependent RNA polymerase located on the DNA of Zea mays chloroplasts has been characterized with respect to its position on the chloroplast genome and its nucleotide sequence. The amino acid sequence derived for a 39 Kd polypeptide shows strong homology with sequences derived from the rpoA genes of other chloroplast species and with the amino acid sequence of the alpha-subunit from E. coli RNA polymerase. Transcripts of the rpoA gene were identified by Northern hybridization and characterized by S1 mapping using total RNA isolated from maize chloroplasts. Antibodies raised against a synthetic C-terminal heptapeptide show cross reactivity with a 39 Kd polypeptide contained in the stroma fraction of maize chloroplasts. It is concluded that the rpoA gene is a functional gene and that therefore, at least the alpha-subunit of plastidic RNA polymerase, is expressed in chloroplasts.
对位于玉米叶绿体DNA上编码依赖DNA的RNA聚合酶α亚基的rpoA基因，在叶绿体基因组上的位置及其核苷酸序列进行了表征。推导的39 Kd多肽的氨基酸序列与其他叶绿体物种rpoA基因推导的序列以及大肠杆菌RNA聚合酶α亚基的氨基酸序列具有很强的同源性。通过Northern杂交鉴定rpoA基因的转录本，并使用从玉米叶绿体中分离的总RNA通过S1作图进行表征。针对合成C末端七肽产生的抗体与玉米叶绿体基质部分中包含的39 Kd多肽显示出交叉反应性。得出的结论是，rpoA基因是一个功能基因，因此，至少质体RNA聚合酶的α亚基在叶绿体中表达。
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