Reference Article: \"High-level expression of bioactive recombinant human lysozy

Question

Reference Article: "High-level expression of bioactive recombinant human lysozyme in the milk of transgenic mice using a modified human lactoferrin BAC" by Shen liu et al.

This article describes a technique for successful strong expression of a human lysozyme gene in mouse mammary gland cells by production of transgenic mice. The journal articles assumes some information is obvious and doesn’t describe the creation of the transgenic mice in complete detail. In a figure, show the steps that produced stable transgenic mice with the hLZ-containing BAC. Then explain the ‘position effect’ (a genetic term) that concerned the researchers before this experiment and the evidence they present that supports the conclusion they have effectively avoided position effect

Explanation / Answer

To express human lysozyme =

First is the need of a candidate vector - which is called as hlF BAC gene - This target gene is replaced by new fragment of gene with the help of plasmids - The gene replacement is exactly done at the Translocation part - replacement done from start to stop codon by flanking regions of gene - a 90 kb 5' & 30 kb 3' - This flanking regions are used for hlZ gene to express - After this position effect - the recombinant new gene was produced rhlZ gene - This was identified and analysed by PCR / Southern blots/Eliza for perfect production of Transgenic mice with rhlZ gene expression - Transgenic mice able to express the antibacterial activity in their milk like native hlZ

The position effect - By changing the location of genes in the chromosomes.This may differ from individual to individual.The 4 Transgenic mice were showing the expression of hlZ gene, which sj=hows antibacterila activity and active components present.

rhLZ was highly expressed in the milk of four transgenic mouse lines (1.20-1.76 g/L), was expressed at a lower level in one additional line (0.21 g/L).This lead to the larger production this hlZ gene in livestocks.

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