Gelvin S, Heizmann P, Howell S H
Proc Natl Acad Sci U S A. 1977 Aug;74(8):3193-7. doi: 10.1073/pnas.74.8.3193.
mRNA coding for the large subunit (LS) of ribulose-1,5-bisphosphate carboxylase [3-phospho-D-glycerate carboxy-lyase (dimerizing), EC 4.1.1.39] from Chlamydomonas reinhardi has been isolated from small polyribosomes immunoadsorbed to column-bound anti-LS antibody. 32P-Labeled LS mRNA was used as a hybridization probe to detect LS genes. The probe hybridized to C. reinhardi chloroplast DNA and at hybridization saturation revealed that there are approximately 75 LS genes per chloroplast. When chloroplast DNA was digested with the restriction endonuclease EcoRI and the fragments were transferred to a nitrocellulose filter, the LS mRNA probe hybridized to a DNA fragment of molecular weight 3.2 X 10(6). This same fragment codes (in part) for 16S and 23S chloroplast rRNAs, which are also coded (in part) by fragments of molecular weights 9.0, 2.3, and 0.4 X 10(6). The restriction fragment containing the LS gene has been cloned in the Escherichia coli plasmid pMB9.
编码莱茵衣藻1,5 - 二磷酸核酮糖羧化酶[3 - 磷酸 - D - 甘油酸羧基裂解酶（二聚化），EC 4.1.1.39]大亚基（LS）的信使核糖核酸（mRNA）已从小型多核糖体中分离出来，这些多核糖体通过免疫吸附在柱结合的抗LS抗体上。用32P标记的LS mRNA作为杂交探针来检测LS基因。该探针与莱茵衣藻叶绿体DNA杂交，在杂交饱和时显示每个叶绿体约有75个LS基因。当用限制性内切酶EcoRI消化叶绿体DNA并将片段转移到硝酸纤维素滤膜上时，LS mRNA探针与分子量为3.2×10^6的DNA片段杂交。这个相同的片段（部分）编码16S和23S叶绿体核糖体RNA，它们也（部分）由分子量为9.0、2.3和0.4×10^6的片段编码。含有LS基因的限制性片段已克隆到大肠杆菌质粒pMB9中。
