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Use Of Polymerase Chain Reaction (PCR) | Production of Vitamins

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Use of Polymerase Chain Reaction ( PCR )

  • PCR is a technique by which any piece of DNA can be quickly amplified ( copied many times ) without using cells.
  • The DNA is incubated in a test tube with a special kind of DNA polymerase enzyme, a mixture of deoxyribonuleotides for use as raw material, and short pieces of synthetic single stranded DNA to serve as primers for DNA synthesis.
  • PCR can make billions of copies of a DNA segment in a few hours.
  • To clone a DNA segment in bacteria takes days.
  • When the source of DNA is scanty or impure, it can be amplified by PCR technique.
  • Amplification can make the identification of DNA easier.
  • Since the sequence of HIV DNA is known, its amplification by PCR can help detect HIV DNA in blood or tissue samples.
  • This is often the best way to detect infection.
  • DNA from single embryonic cell is amplified by PCR for rapid prenatal diagnosis of genetic disorders.
  • DNA from tiny amounts of blood, tissue or semen found at the site of crime can be amplified to facilitate its iden tification.
  • DNA technology can help identify individuals with genetic disorders before the appearance of symptoms, even before birth.
  • It is also possible to identify symptomless carriers of potentially harmful recessive alleles, such as of haemophilia, phenylketonurea (PKU).
  • PCR is also helpful in DNA fingerprinting.
Production of Vitamins 
  • Vitamin B2 ( Riboflavin )
  • Riboflavin is produced commercially by direct fermenta tion utilizing the fungus Ashbya gossypii.
  • Fermentation conditions -pH 6.0 to 7.5.4 to 5 days at 28-30 ° C, aerobic.
  • Vitamin B12 ( Cobalamine )
  • Now a days vitamin B12 is produced by a direct fermentation utilizing bacteria such as Propionibacterium shermanii, P. frendenreichii and Pseudomonas denitrificans.
  • Fermentation conditions -pH 6 to 7.7 . 2 days at 26-28 ° C. aerobic
  • Vitamin C ( Ascorbic Acid )
  • Vitamin C was first recognized when in 1747, Scottish naval surgeon James Lind discovered that something in Citrus foods prevented scurvy.
  • In 1928 Albert Szent - Gyorgyi, a much admired biochemist, was the first to isolate vitamin C ( ascorbic acid ).
  • He later won the 1937 Nobel Prize in Physiology and Medicine for other work.
  • Vitamin C was the first vitamin to be synthesized artificially in a process invented by Dr. Tadeusz Reichstein, of the Swiss Institute of Technology in Zurich in 1939.
  • Vitamin C is produced by utilizing Gluconobacter oxydans.
  • Fermentation conditions -pH 7, 45 hours at 30 ° C, aerobic.
  • Vitamin A ( B - Carotene )
  • B - carotene is produced by members of choanephoraceae family of phycomycetes . Phycomyces blakesleeanus, Choanephora cucurbitarum and Blackeslea trispora have been extensively studied for their ability to produce B - carotene.
Making a Choice of Baby's Sex
  • Recently techniques have also been developed which will not require preferential abor tion but will allow preferential fertilization by male (carrying Y chromosome) or female (carrying X chromosome) determining sperms.
  • There are techniques available now, which allow separation of sperms carrying Y chromosomes, from the ejaculate of a man ( through Ericson's method developed by R. Ericson of U.S.A ) to be used for insemination of ovulating women.
  • This technique (using quinacrine stain) has been used with 80 % success in 47 sperm centres in the world including one in Mumbai.
  • Ericson has actually established a company named Gametrics Ltd, in California , U.S.A. which specializes in separating sperms with Y chromosome and hundreds of male children have been produced with its help.
  • Techniques have also been developed to separate sperms carrying X chromosome for artificial insemination leading to the birth of female children.
  • This technique involves ' sephadex gel column ' in which sperms with Y, being lighter are trapped in gel and those with X being heavier reach the bottom of the column, and can be used for inseminaton.

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