Biotech & Its Applications
Genetically modified microbes, fungi, plants, animals ko use karke industrial scale par biopharmaceuticals aur biologicals produce karna.
1. Three Critical Research Areas
- Best catalyst (microbe/enzyme) provide karna.
- Engineering ke through catalyst ke liye optimal conditions create karna.
- Downstream processing technologies (protein purification ke liye).
2. Applications in Agriculture
Food production badhane ke 3 options hain:
- Agro-chemical based (Fertilizers/Pesticides).
- Organic agriculture.
- Genetically engineered crop-based (Best option).
Green Revolution ne yield 3x ki thi, par fir bhi growing population ke liye enough nahi thi (Agrochemicals mehnge hote hain).
GMOs (Genetically Modified Organisms)
Plants, bacteria, fungi aur animals jinke genes alter kiye gaye hain. GM Plants ke fayde:
- Abiotic stress (cold, drought, salt) ke liye zyada tolerant.
- Pest-resistant crops (chemical pesticides par reliance kam).
- Post-harvest losses ko kam karna.
- Mineral usage ki efficiency badhana (soil jaldi exhaust nahi hoti).
- VVI Nutritional value badhana e.g., Golden Rice (Vitamin A enriched).
3. Bt Cotton (Pest Resistance)
Bt = Bacillus thuringiensis (Ek bacterium).
Bt toxin gene bacterium se clone karke plants (cotton, corn, rice) me daal diya gaya hai. Isse plant khud insects ko maar sakta hai bina insecticides ke (Bio-pesticide).
Mechanism of Bt Toxin:
- Bt bacteria special protein crystals banate hain jisme toxic insecticidal protein hota hai.
- Ques: Toxin bacteria ko kyu nahi marta?
Ans: Kyunki bacteria me ye inactive protoxin ke form me hota hai. - Jaise hi insect ise khata hai, uske gut ke Alkaline pH ke karan protoxin active toxin me convert ho jata hai.
- Active toxin midgut epithelial cells se bind karke pores (chhed) banata hai -> cell swelling -> lysis -> insect ki death.
RATTA MAAR GENES! Specificity
Bt toxins insect-group specific hote hain. Gene ko cry kehte hain.
- cryIAc & cryIIAb : Cotton bollworms ko control karte hain.
- cryIAb : Corn borer ko control karta hai.
4. Pest Resistant Plants (Nematodes)
Name Meloidogyne incognita (ek nematode) tobacco plants ki roots ko infect karke yield bohat kam kar deta hai.
Isse bachne ke liye RNA interference (RNAi) strategy use ki gayi.
RNA interference (RNAi):
- Ye sabhi eukaryotic organisms me cellular defense ka ek natural method hai.
- Isme specific mRNA ko "silence" kar diya jata hai using complementary dsRNA (double-stranded RNA).
- dsRNA bind karke mRNA ki translation rok deta hai (silencing).
- Vector used: Agrobacterium vectors use karke nematode-specific genes host plant me daale gaye.
- Aise daala gaya ki plant dono sense aur anti-sense RNA produce kare, jo milkar dsRNA banate hain aur RNAi initiate karte hain. Parasite survive nahi kar pata!
5. Biotech in Medicine
Recombinant DNA tech ne safe & effective therapeutic drugs banaye hain (No unwanted immunological responses jo non-human sources se aati thi).
- Data World me 30 recombinant therapeutics approved hain.
- Data India me inme se 12 presently market ho rahi hain.
6. Genetically Engineered Insulin
Diabetes ke liye use hota hai. Pehle insulin slaughtered pigs & cattle ke pancreas se nikalte the.
Problem: Kuch patients me animal insulin se allergies aur side-effects hote the.
Insulin Structure:
- 2 short polypeptide chains hoti hain: Chain A & Chain B.
- Dono chains ek dusre se Disulphide bridges se linked hoti hain.
- Mammals me insulin ek pro-hormone (Pro-insulin) ki tarah synthesize hota hai jisme ek extra stretch hota hai called C-peptide.
- Mature aur functional insulin me C-peptide nahi hota (maturation ke time remove ho jata hai).
VVI Year & Company
1983 me Eli Lilly (American company) ne A & B chains ke DNA sequences banaye aur unhe E. coli plasmids me daal kar alag-alag produce kiya. Baad me dono ko nikal kar disulphide bonds banaye = Human Insulin (Humulin).
7. Gene Therapy
Genetic defects ko embryo ya child stage me hi theek karna. Ek normal gene ko individual/embryo me deliver kiya jata hai defective gene ko compensate karne ke liye.
First Case! ADA Deficiency
- First clinical gene therapy 1990 me ek 4-year old girl ko di gayi thi.
- Disease: ADA (Adenosine deaminase) deficiency.
- ADA enzyme immune system ke function ke liye crucial hota hai. Ye disorder gene ke deletion se hota hai.
Treatments for ADA:
- Traditional: Bone marrow transplantation ya Enzyme Replacement Therapy (Injection of ADA). Lekin ye dono completely curative nahi hain.
- Gene Therapy Method:
- Patient ke blood se lymphocytes culture karte hain.
- Ek retroviral vector use karke functional ADA cDNA (m-RNA se bana hua) lymphocytes me daalte hain.
- In cells ko wapas patient me daal dete hain.
- Problem: Lymphocytes immortal nahi hain, toh patient ko periodic infusions chahiye hote hain.
- Permanent Cure: Agar bone marrow cells se gene nikal kar early embryonic stages me introduce kar dein.
8. Molecular Diagnosis
Serum/Urine analysis jaise traditional methods se early detection possible nahi hai. Early detection ke liye 3 methods hain:
- PCR (Polymerase Chain Reaction): Jab bacteria/virus body me bohat kam hote hain (symptoms nahi aate), PCR unke nucleic acid ko amplify kar deta hai.
Uses: Suspected AIDS patients me HIV detect karna, Cancer patients me gene mutations detect karna. - ELISA: (Enzyme Linked Immunosorbent Assay). Ye Antigen-Antibody interaction ke principle par based hai. Ya toh pathogen (antigen) detect karo ya antibodies detect karo.
- Autoradiography: ssDNA/RNA jisme radioactive molecule (probe) laga ho, wo mutated gene se bind nahi karta aur photographic film par appear nahi hota.
9. Transgenic Animals
Wo animals jinka DNA manipulate kiya gaya hai ek extra (foreign) gene express karne ke liye.
Data: 95% se zyada transgenic animals Mice hain.
Kyu banate hain inko?
- Normal physiology & development: Genes ka body par kya asar hota hai study karne ke liye (e.g., Insulin-like growth factor).
- Study of disease: Cancer, Cystic fibrosis, Rheumatoid arthritis, Alzheimer's ke models banaye jate hain.
More Reasons for Transgenic Animals:
- Biological Products: Mahangi dawaiyan banan e ke liye.
- Name Alpha-1-antitrypsin banaya gaya to treat Emphysema. (Phenylketonuria/Cystic fibrosis ke attempts bhi hue hain).
- Rosie (1997): First transgenic cow. Iske milk me human protein tha (2.4 gm/litre). Milk me human alpha-lactalbumin tha, jo human babies ke liye normal cow milk se zyada balanced hai.
- Vaccine Safety: Transgenic mice use hote hain Polio vaccine safety test karne ke liye (Monkeys replace karne ke liye).
- Chemical Safety Testing: Toxicity testing, mice ko zyada sensitive banaya jata hai toxic substances se.