Silverthorne J, Ellis R J
Biochim Biophys Acta. 1980 Apr 30;607(2):319-30. doi: 10.1016/0005-2787(80)90084-2.
Excised primary leaves of spinach (Spinacia oleracea) incorporate [35S]-methionine into a number of chloroplast polypeptides. The ratio of incorporation of isotope into the large subunit of ribulose bisphosphate carboxylase relative to a thylakoid polypeptide (peak D) decreases during leaf development in whole leaves; this changing pattern of incorporation is also observed in isolated chloroplasts where these two polypeptides are the major products of protein synthesis. Chloroplast RNA prepared from developing leaves was translated in a reticulocyte lysate extract to yield full-length carboxylase large subunit and peak D polypeptides. The fidelity of translation of these two polypeptides was checked by partial protease digestion. Changes in the synthesis of the large subunit of the carboxylase and peak D in developing leaves are reflected in changes in the amount of translatable mRNA for these two polypeptides.
菠菜（Spinacia oleracea）的离体初生叶能将[35S] - 甲硫氨酸掺入多种叶绿体多肽中。在全叶的叶片发育过程中，相对于类囊体多肽（峰D），同位素掺入核酮糖二磷酸羧化酶大亚基的比例会降低；在分离的叶绿体中也观察到这种变化的掺入模式，其中这两种多肽是蛋白质合成的主要产物。从发育中的叶片制备的叶绿体RNA在网织红细胞裂解物提取物中进行翻译，以产生全长羧化酶大亚基和峰D多肽。通过部分蛋白酶消化检查这两种多肽翻译的保真度。发育中叶片中羧化酶大亚基和峰D合成的变化反映在这两种多肽可翻译mRNA量的变化上。
