Hess J F, Casselman J T, FitzGerald P G
Department of Cell Biology and Human Anatomy, School of Medicine, University of California, Davis 95616, USA.
Curr Eye Res. 1995 Jan;14(1):11-8. doi: 10.3109/02713689508999909.
We have used the polymerase chain reaction (PCR) to amplify CP 115 and CP 47 encoding sequences from human lens cDNA samples. DNA sequence and northern blot analysis were used to confirm human origin. From the determined cDNA sequences, human-specific oligonucleotides were synthesized and assessed for the ability to amplify human genomic DNA. After empirically selecting a primer pair for each gene able to amplify human genomic DNA, and optimizing PCR conditions for human specificity, we used the PCR to screen a panel of mouse/human somatic cell hybrid DNA samples. Amplification of CP 115 or CP 47 sequences in each of the somatic cell hybrid samples was correlated with the presence/absence of human genomic DNA sequences encoding the respective gene sequences. From our results, we conclude that the gene for human CP 115 resides on chromosome 20 and the gene for human CP 47 on chromosome 3. Further mapping using somatic cell lines carrying derivatives of human chromosome 3 localize the gene for CP 47 to 3q21-25. We propose LIFL-H (Lens Intermediate Filament Like-Heavy) for CP 115 and LIFL-L (Lens Intermediate Filament Like-Light) for CP 47 as the gene symbols for these loci.
我们利用聚合酶链反应(PCR)从人晶状体cDNA样本中扩增CP 115和CP 47的编码序列。采用DNA序列分析和Northern印迹分析来确认其人类来源。根据测定的cDNA序列,合成了人特异性寡核苷酸,并评估其扩增人基因组DNA的能力。在凭经验为每个基因选择了一对能够扩增人基因组DNA的引物,并优化了针对人特异性的PCR条件后,我们使用PCR筛选了一组小鼠/人类体细胞杂交DNA样本。每个体细胞杂交样本中CP 115或CP 47序列的扩增与编码相应基因序列的人基因组DNA序列的有无相关。根据我们的结果,我们得出结论,人CP 115基因位于20号染色体上,人CP 47基因位于3号染色体上。使用携带人3号染色体衍生物的体细胞系进行进一步定位,将CP 47基因定位到3q21 - 25。我们提议将CP 115的基因符号命名为LIFL - H(晶状体中间丝样重链),将CP 47的基因符号命名为LIFL - L(晶状体中间丝样轻链)。