Dolly was the first mammal (Sheep) clone developed by Ian Wilmut and Campbell in 1997. Dolly, the transgenic clone was developed by the nuclear transfer technique and the phenomenon of totipotency. Totipotency refers to the potential of a cell to develop different cells, tissues, organs and finally an organism.
The mammary gland udder cells (somatic cells) from a donor sheep (ewe) were isolated and subjected to starvation for 5 days. The udder cells could not undergo normal growth cycle, entered a dormant stage and became totipotent. An ovum (egg cell) was taken from another sheep (ewe) and its nucleus was removed to form an enucleated ovum. The dormant mammary gland cell/udder cell and the enucleated ovum were fused. The outer membrane of the mammary cell was ruptured allowing the ovum to envelope the nucleus.
The fused cell was implanted into another ewe which served as a surrogate mother. Five months later dolly was born. Dolly was the first animal to be cloned from a differentiated somatic cell taken from an adult animal without the process of fertilization.

The various steps involved in the production of transgenic organisms are,
(i) Identification and separation of desired gene.
(ii) Selection of a vector (generally a virus) or direct transmission.
(iii) Combining the desired gene with the vector.
(iv) Introduction of transferred vector into cells, tissues, embryo or mature individual.
(v) Demonstration of integration and expression of foreign gene in transgenic tissue or animals. Transgenic animals such as mice, rat, rabbit, pig, cow, goat, sheep and fish have been produced.

Stem cells are undifferentiated cells found in most of the multi cellular animals. These cells maintain their undifferentiated state even after undergoing numerous mitotic divisions. Stem cell research has the potential to revolutionize the future of medicine with the ability to regenerate damaged and diseased organs. Stem cells are capable of self renewal and exhibit 'cellular potency'. Stem cells can differentiate into all types of cells that are derived from any of the three germ layers ectoderm, endoderm and mesoderm.
In mammals there are two main types of stem cells - embryonic stem cells (ES cells) and adult stem cells. ES cells are pluripotent and can produce the three primary germ layers ectoderm, mesoderm and endoderm. Embryonic stem cells are multipotent stem cells that can differentiate into a number of types of cells. ES cells are isolated from the epiblast tissue of the inner cell mass of a blastocyst. When stimulated ES can develop into more than 200 cells types of the adult body. ES cells are immortal ie. they can proliferate in a sterile culture medium and maintain their undifferentiated state. Adult stem cells are found in various tissues of children as well as adults. An adult stem cell or somatic stem cell can divide and create another cell similar to it. Most of the adult stem cells are multipotent and can act as a repair system of the body, replenishing adult tissues. The red bone marrow is a rich source of adult stem cells.
The most important and potential application of human stem cells is the generation of cells and tissues that could be used for cell based therapies. Human stem cells could be used to test new drugs.

(i) Dolly was the first mammal (Sheep) clone developed by Ian Wilmut and Campbell in 1997. Dolly, the transgenic clone was developed by the nuclear transfer technique and the phenomenon of totipotency. Totipotency refers to the potential of a cell to develop different cells, tissues, organs and finally an organism

(ii) The mammary gland udder cells (somatic cells) from a donor sheep (ewe) were isolated and subjected to starvation for 5 days. The udder cells could not undergo normal growth cycle, entered a dormant stage and became totipotent. An ovum (egg cell) was taken from another sheep (ewe)) and its nucleus was removed to form an enucleated ovum. The dormant mammary gland cell/udder cell and the enucleated ovum were fused. The outer membrane of the mammary cell was ruptured allowing the ovum to envelope the nucleus. The fused cell was implanted into another ewe which served as a surrogate mother. Five months later dolly was born. Dolly was the first animal to be cloned from a differentiated somatic cell taken from an adult animal without the process.

1. Production of insulin by recombinant DNA technology started in the late 1970s.
2. This technique involved in the insertion of human insulin gene on the plasmids of E. Coli. The polypeptide chains are synthesized as a precursor called pre-proinsulin as it contains A and B segments linked by a third chain (c) and preceded by a leader sequence.
3. The leader sequence is removed after translation and 'c' chain is excised, leaving the A and B polypeptide chains.
4. Insulin was the first ever pharmaceutical product of r-DNA, technology.