Nichols B J, Hall L, Perry A C, Denton R M
Department of Biochemistry, School of Medical Sciences, University of Bristol, U.K.
Biochem J. 1993 Oct 15;295 ( Pt 2)(Pt 2):347-50. doi: 10.1042/bj2950347.
A 600 bp cDNA fragment encoding part of the gamma-subunit of pig heart NAD(+)-isocitrate dehydrogenase (ICDH gamma) was amplified by PCR using redundant oligonucleotide primers based on partial peptide sequence data [Huang and Colman (1990) Biochemistry 29, 8266-8273]. This PCR fragment was then used as a probe to isolate clones encoding the complete mature forms of the gamma-subunit from rat epididymis and monkey testis cDNA libraries. Comparison of the deduced amino acid sequences of the rat and monkey subunits and the partial sequence of the pig heart enzyme revealed a remarkably high level of sequence identity. The relationship between the deduced amino acid sequences of the NAD(+)-ICDH gamma-subunits and those of nonmammalian NAD(+)- and NADP(+)-ICDH subunits is discussed.
利用基于部分肽序列数据设计的简并寡核苷酸引物,通过聚合酶链式反应(PCR)扩增出一段600 bp的cDNA片段,该片段编码猪心脏NAD(+)-异柠檬酸脱氢酶(ICDHγ)γ亚基的部分序列[Huang和Colman(1990年),《生物化学》29卷,8266 - 8273页]。然后将该PCR片段用作探针,从大鼠附睾和猴睾丸cDNA文库中分离出编码γ亚基完整成熟形式的克隆。大鼠和猴亚基推导的氨基酸序列与猪心脏酶的部分序列比较显示,序列同一性水平非常高。文中讨论了NAD(+)-ICDHγ亚基推导的氨基酸序列与非哺乳动物NAD(+)-和NADP(+)-ICDH亚基推导的氨基酸序列之间的关系。