How are genes measured? Examples from studies on iron metabolism in pregnancy.
As the 21st century moves forward, it is becoming more and more apparent that the genetic makeup of any individual strongly influences the way they metabolise nutrients. It is very important, therefore, to understand the techniques and technologies used to assess the contribution genes make to the physiology of an individual. Clearly, it is not possible to provide a comprehensive overview, but in the present review an attempt will be made to show, using examples from the authors' research, how these methods have contributed to this understanding. Studies are being undertaken into Fe transport across the placenta, from the mother to the fetus, and the consequences of maternal anaemia on pregnancy outcome. Levels of gene transcript and protein have been measured using Northern and Western blotting respectively. During the course of this work a new protein has been identified using the available human genome database. Following this 'in silico' or 'cyber biology', techniques such as real-time RT-PCR and RNA interference have been used to examine expression of this gene and its protein. The methods used, briefly how they work and some of their limitations will be explained. The objective of the present review is primarily to give a better perception of how molecular biology can be used in research and to help gain a clearer understanding of some of the techniques used.
Andersen
Henriette S
HS
Development, Growth and Function Division, Rowett Research Institute, Bucksburn, Aberdeen AB21 9SB, UK. H.Andersen@rowett.ac.uk
McArdle
Harry J
HJ
D018798:Anemia, Iron-Deficiency / Q000235:genetics / Q000378:metabolism* / Q000503:physiopathology; D015152:Blotting, Northern; D015153:Blotting, Western; D003300:Copper / Q000378:metabolism; D005314:Embryonic and Fetal Development; D005260:Female; D014644:Genetic Variation; D006801:Humans; D007501:Iron / Q000378:metabolism*; D000090463:Iron Deficiencies; D008431:Maternal-Fetal Exchange / Q000502:physiology; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism*; D010920:Placenta / Q000378:metabolism; D011247:Pregnancy / Q000378:metabolism*; D011256:Pregnancy Outcome; D020133:Reverse Transcriptase Polymerase Chain Reaction
D003300:Copper; D007501:Iron; D010088:Oxidoreductases